CN113966345A

CN113966345A - Natriuretic peptide receptor 1 antibodies and methods of use

Info

Publication number: CN113966345A
Application number: CN202080043331.7A
Authority: CN
Inventors: J·L·迪纳; L·加特克; F·哈特勒普; T·胡; K·拉德茨基-贝斯; M·罗曼诺夫斯基; C·鲁索; X·维兹勒; X·谢
Original assignee: Novartis AG
Current assignee: Novartis AG
Priority date: 2019-06-12
Filing date: 2020-06-10
Publication date: 2022-01-21
Also published as: AR119267A1; CO2021016619A2; CA3141561A1; US20200392225A1; MA56180A; CR20210607A; IL288143A; KR20220019785A; CL2021003266A1; WO2020250159A1; ECSP21088669A; AU2020291821A1; US20220356242A1; MX2021015160A; US20230365674A1; CU20210099A7; TW202112816A; EP3983437A1; UY38747A; EA202193345A1

Abstract

The present disclosure relates to anti-natriuretic peptide receptor 1(NPR1) antibodies, including agonist antibodies capable of activating the NPR1 receptor; a pharmaceutical composition comprising the antibody; and methods of treatment comprising the antibodies.

Description

Natriuretic peptide receptor 1 antibodies and methods of use

Background

Heart failure is a significant public health problem involving over 2000 million patients worldwide (Orso et al 2014, Expert Opin Pharmacother [ Pharmacology experts ]15(13): 1849-. Natriuretic Peptide Receptor 1 (natural Peptide Receptor 1, NPR 1; also known as NPRA) is a Receptor guanylate cyclase that is activated by Atrial Natriuretic Peptide (ANP) resulting in a decrease in blood pressure and blood volume (Chen & Burnett,2006, European Heart Journal supplement 8 (supplement E): E18-E25; ibeburoku et al 2011, supra; ari et al 2015, Bioscience Reports 35(5): E00260). ANP binding induces dimerization and twisting of The receptor, thereby inducing activation of The guanylate cyclase domain and conversion of GTP to cGMP (Misono et al, 2011, The FEBS journal [ FEBS J ]278(11):1818 and 1829). ANP is cleared by NPR3, a natriuretic peptide receptor lacking the guanylate cyclase domain, and degraded by Neutral Endopeptidase (NEP) (Chen & Burnett 2006, supra; Schmitt et al 2003, Clin Sci (Lond) [ clinical sciences (London) ]105(2): 141-. Certain antibodies (including antibody 5591-IgG) against NPR1 are described, for example, in WO 2010/065293. However, these antibodies appear to be functionally inactive in vitro in the absence of ANP, and not functionally active in vivo.

It has been shown that an increase in ANP may be beneficial for patients with heart failure (heart pumping outward) with reduced ejection fraction. See McMurray et al, n.engl.j.med. [ new england journal of medicine ]; 371 th volume, 11 th stage, 993 th and 1004 th pages (2014); and nougue et al, Eur J Heart Fail. [ european journal of Heart failure ]2019, month 5; 21(5):598-605. However, there is a need for agents with longer acting periods that have alternative modes of action to supplement or replace existing therapies.

Disclosure of Invention

It is demonstrated herein that NPR1 can be activated by using an agonistic anti-NPR 1 antibody or antigen binding fragment thereof. Furthermore, the present disclosure demonstrates that there are two types of such antibodies. One type binds to NPR1 and competes with ANP binding (but still activates NPR 1; hereinafter referred to as "ANP-competitive" anti-NPR 1 antibody), while the second type is capable of binding to and activating NPR1 without competing with ANP (hereinafter referred to as "ANP-noncompetitive" anti-NPR 1 antibody). Such antibodies (e.g., ANP non-competitive anti-NPR 1 antibodies) can be used to enhance the body's natural system and/or to consolidate existing therapeutic principles. Furthermore, it has been found that certain NPR1 agonist antibodies capable of activating NPR1 in the absence of ANP are functionally equivalent to ANP.

The antibodies of the present application show in vivo activity in both mice and rats. In addition, unique epitope binding of the antibodies described herein has been demonstrated using crystal structure data.

Accordingly, the present disclosure provides anti-NPR 1 antibodies (e.g., human monoclonal antibodies) or antigen-binding fragments thereof that (i) bind to natriuretic peptide receptor 1(NPR 1); and (ii) is capable of activating NPR1 in the absence of ANP. Such antibodies are agonistic anti-NPR 1 antibodies. In some embodiments of the invention, the disclosure also provides an anti-NPR 1 antibody, or antigen-binding fragment thereof, that (i) binds to natriuretic peptide receptor 1(NPR 1); and (ii) activates NPR1 in the absence of ANP. In some embodiments of the invention, the disclosure also provides an antibody or antigen-binding fragment thereof that (i) binds to natriuretic peptide receptor 1(NPR 1); and (ii) is capable of activating NPR1 in both the presence and absence of ANP. Also provided are nucleic acids encoding the antibodies, vectors comprising the nucleic acids, host cells comprising the nucleic acids and/or vectors, and methods of making the antibodies using the nucleic acids, vectors, and/or host cells. Also provided are pharmaceutical compositions and combinations comprising the antibodies, nucleic acids, vectors, or host cells, and methods of treatment using the antibodies, nucleic acids, vectors, host cells, or pharmaceutical compositions. Also disclosed herein is the use of the antibody, nucleic acid, vector, host cell or pharmaceutical composition or combination in the treatment of a disease.

Accordingly, in one aspect of the invention, provided herein is an isolated antibody or antigen-binding fragment that (i) binds to natriuretic peptide receptor 1(NPR 1); and (ii) is capable of activating NPR1 in the absence of Atrial Natriuretic Peptide (ANP). In some embodiments of the invention, the isolated antibody or antigen binding fragment does not bind to and/or does not activate natriuretic peptide receptor 2(NPR2) and/or natriuretic peptide receptor 3(NPR 3). In some embodiments of the invention, the isolated antibody or antigen-binding fragment binds to (a) human NPR 1; and (b) mouse NPR1 and/or rat NPR 1.

In some embodiments of the invention, the antibody or antigen-binding fragment binds to (a) human NPR 1; and (b) cynomolgus monkey NPR 1. In some embodiments of the invention, the antibody or antigen binding fragment is ANP non-competitive. In some embodiments of the invention, the antibody or antigen binding fragment is ANP competitive. In some embodiments of the invention, the antibody or antigen binding fragment is capable of stabilizing the ANP-NPR1 complex.

In some embodiments of the invention, the antibody or antigen binding fragment binds to an epitope within amino acids 99-133 of SEQ ID NO. 1, e.g., within the region of human NPR1 encompassed by amino acids 99-133 of SEQ ID NO. 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to an epitope comprising at least two amino acid residues within amino acids 99-133 of SEQ ID NO: 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to an epitope comprising at least 3, 4, 5, 6, 7, or 8 amino acid residues within amino acids 99-133 of SEQ ID NO: 1.

In some embodiments of the invention, the antibody or antigen binding fragment binds to an epitope within amino acids 99-111 of SEQ ID NO. 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to an epitope within amino acids 99-103 of SEQ ID NO. 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to an epitope within amino acids 105 and 111 of SEQ ID NO: 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to an epitope comprising at least 2, 3, or 4 amino acid residues within amino acid 105-111 of SEQ ID NO 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to a conformational epitope of human NPR1, and wherein the conformational epitope comprises at least one amino acid residue each of (i) amino acids 99-103 of SEQ ID NO:1, (ii) amino acids 105-111 of SEQ ID NO:1, (iii) amino acids 131-134 of SEQ ID NO:1, and further binds to amino acids 375 and/or 378 of SEQ ID NO: 1. In some embodiments of the invention, the epitope is a conformational epitope, and the conformational epitope additionally comprises at least one amino acid residue selected from the group consisting of amino acids 33, 34, 76, 82, and 104 of SEQ ID No. 1. In some embodiments of the invention, the conformational epitope additionally comprises at least one amino acid residue selected from the group consisting of amino acids 33, 34, 76, 82, 104, 374 and 375 of SEQ ID No. 1.

In some embodiments of the invention, the antibody or antigen binding fragment binds to at least

amino acids

82, 102, 103, 105, 106, 109, 132, and 375 of SEQ ID NO. 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to at least

amino acids

34, 82, 102, 103, 105, 106, 107, 109, 132, 133, 375, and 378 of SEQ ID No. 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to at least

amino acids

79, 82, 99, 102, 103, 105, 106, 109, 131, 132, and 375 of SEQ ID No. 1.

In some embodiments of the invention, the antibody or antigen binding fragment binds to an epitope within amino acids 188-219 of SEQ ID NO: 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to an epitope comprising at least 2, 3, 4, 5, 6, or 7 amino acids within amino acids 188-219 of SEQ ID NO: 1. In some embodiments of the invention, the antibody or antigen binding fragment binds to a conformational epitope within NPR1, and the conformational epitope comprises at least one amino acid residue in each of (i) amino acids 188-198 of SEQ ID NO:1, (ii) amino acids 201-208 of SEQ ID NO:1, (iii) amino acids 215-238 of SEQ ID NO:1, and (iv) amino acids 294-297 of SEQ ID NO: 1. In some embodiments of the invention, the antibody or antigen-binding fragment binds to at least amino acids 188, 192, 194, 197, 201, 208, and 219 of SEQ ID No. 1. In some embodiments of the invention, the antibody or antigen-binding fragment binds to at least amino acids 188, 192, 194, 197, 201, 208, 219, and 295 of SEQ ID No. 1.

In some embodiments of the invention, the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3), wherein: (a) (I) HCDR1Comprises or consists of the amino acid sequence shown as SEQ ID NO. 28; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆YADSVKG (SEQ ID NO:429) or consists of the amino acid sequence shown in YADSVKG (SEQ ID NO:429), wherein X₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X₆Is Y or F; HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30; LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41; LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 42; and LCDR3 contains, for example, Y₁QY₂Y₃Y₄Y₅An amino acid sequence shown by PRT (SEQ ID NO:430) or consists of the amino acid sequence; wherein Y is₁Is M or Q, Y₂Is S, E, T or I, Y₃Is Y or W, Y₄Is E, V, R, A, T or M, and Y₅Is K, V, R or A; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 31; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆YADSVKG (SEQ ID NO:429) or consists of the amino acid sequence shown in YADSVKG (SEQ ID NO:429), wherein X₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X ₆Is Y or F; the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 42, and the LCDR3 comprises or consists of Y₁QY₂Y₃Y₄Y₅An amino acid sequence shown by PRT (SEQ ID NO:430) or consists of the amino acid sequence; wherein Y is₁Is M or Q, Y₂Is S, E, T or I, Y₃Is Y or W, Y₄Is E, V, R, A, T or M, and Y₅Is K, V, R or A; (III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:32, HCDR2 comprises₁SX₂GX₃Y (SEQ ID NO:431) or an amino acid sequence consisting thereof, wherein X₁Is S or E, X₂Is D or K, or X₃Is S or N, HCDR3 comprises the sequence shown in SEQ ID NO 30, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 44, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 45, and LCDR3 comprises or consists of Y₁Y₂Y₃Y₄PR (SEQ ID NO:432) or consists of the amino acid sequence thereof; wherein Y is₁Is S, E, T or I, Y₂Is Y or W, Y₃Is E, V, R, A, T or M, and Y₄Is K, V, R or A; or (IV) the HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:34 and the HCDR2 comprises the amino acid sequence shown in IX ₁SX₂GX₃YX₄(SEQ ID NO:433) or consists of the amino acid sequence shown in (wherein X is₁Is S or E, X₂Is D or K, X₃Is S or N, and X₄Is I or T, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:36, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:47, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and LCDR3 comprises or consists of Y₁QY₂Y₃Y₄Y₅An amino acid sequence shown by PRT (SEQ ID NO:430) or consists of the amino acid sequence; wherein Y is₁Is M or Q, Y₂Is S, E, T or I, Y₃Is Y or W, Y₄Is E, V, R, A, T or M, and Y₅Is K, V, R or A; (b) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 28; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆YADSVKG (SEQ ID NO:429) or consists of the amino acid sequence shown in YADSVKG (SEQ ID NO:429), wherein X₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X₆Is Y or F; HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30; LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41; LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 42; and LCDR3 comprises e.g. QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y ₂Is V, R, A, T or M, and Y₃Is K, V, R orA; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 31; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆YADSVKG (SEQ ID NO:429) or consists of the amino acid sequence shown in YADSVKG (SEQ ID NO:429), wherein X₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X₆Is Y or F; the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 42, and the LCDR3 comprises or consists of the amino acid sequence shown as QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; (III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:32, HCDR2 comprises₁SX₂GX₃Y (SEQ ID NO:431) or an amino acid sequence consisting thereof, wherein X₁Is S or E, X₂Is D or K, or X₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 44, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 45, and the LCDR3 comprises or consists of the amino acid sequence shown as Y ₁WY₂Y₃PR (SEQ ID NO:435) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; or (IV) the HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:34 and the HCDR2 comprises the amino acid sequence shown in IX₁SX₂GX₃YX₄(SEQ ID NO:433) or consists of the amino acid sequence shown in (wherein X is₁Is S or E, X₂Is D or K, X₃Is S or N, and X₄Is I or T, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:36, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:47, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and LCDR3 comprises or consists of the amino acid sequence shown as QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; (c) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 28; HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 119; HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30; LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41; LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 42; and LCDR3 comprises e.g. QQY ₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 31; HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 119; the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 42, and the LCDR3 comprises or consists of the amino acid sequence shown as QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; (III) the HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:32, the HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:120, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:44, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and the LCDR3 comprises or consists of the amino acid sequence shown as Y₁WY₂Y₃PR (SEQ ID NO:435) or consists of the amino acid sequence; wherein Y is ₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; or (IV) the HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:34, the HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:121, the HCDR3 comprises the amino acid sequence shown as SEQ ID NO:36Or consists thereof, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:47, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and the LCDR3 comprises or consists of QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; (d) (I) HCDR1 comprises e.g. GFTFX₁An amino acid sequence shown by THYIH (SEQ ID NO:436) or consists of the same, wherein X₁Is N, S or Q, HCDR2 comprises e.g. SIY₁Y₂Y₃GY₄Y₅TY₆YADSVKG (SEQ ID NO:437) or an amino acid sequence consisting of the same, wherein Y₁Is S or G, Y₂Is S or G, Y₃Is S or Q, Y₄Is S, Q or G, Y₅Is S, N or M, and Y₆Is Y or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 17, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 18, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19; (II) HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:7 and HCDR2 comprises or consists of SIY ₁Y₂Y₃GY₄Y₅TY₆YADSVKG (SEQ ID NO:437) or an amino acid sequence consisting of the same, wherein Y₁Is S or G, Y₂Is S or G, Y₃Is S or Q, Y₄Is S, Q or G, Y₅Is S, N or M, and Y₆Is Y or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 17, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 18, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19; (III) HCDR1 comprises e.g. GFTFX₁TH (SEQ ID NO:438) or an amino acid sequence consisting thereof, wherein X₁Is N, S or Q, HCDR2 comprises e.g. Y₁Y₂Y₃GY₄Y₅(SEQ ID NO:439) or an amino acid sequence consisting thereof, wherein Y₁Is S or G, Y₂Is S or G, Y₃Is S or Q, Y₄Is S, Q or G, and Y₅S, N or M, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 20, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 21, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 22; or (IV) HCDR1 comprises, e.g., GFTFX₁An amino acid sequence represented by THY (SEQ ID NO:440) or consisting thereof, wherein X ₁Is N, S or Q, HCDR2 comprises, for example, IY₁Y₂Y₃GY₄Y₅T (SEQ ID NO:441) or an amino acid sequence consisting thereof, wherein Y₁Is S or G, Y₂Is S or G, Y₃Is S or Q, Y₄Is S, Q or G, and Y₅S, N or M, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 12, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 23, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 21, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19; (e) (I) HCDR1 comprises e.g. GFTFX₁An amino acid sequence shown by THYIH (SEQ ID NO:436) or consists of the same, wherein X₁Is N, S or Q, HCDR2 comprises e.g. SISY₁SGY₂Y₃TYYADSVKG (SEQ ID NO:442), wherein Y is₁Is S or G, Y₂Is S or Q, and Y₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 17, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 18, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19; (II) HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:7 and HCDR2 comprises a sequence as SISY ₁SGY₂Y₃TYYADSVKG (SEQ ID NO:442), wherein Y is₁Is S or G, Y₂Is S or Q, and Y₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 17, the LCDR2 comprises18 and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 19; (III) HCDR1 comprises e.g. GFTFX₁TH (SEQ ID NO:438) or an amino acid sequence consisting thereof, wherein X₁Is N, S or Q, HCDR2 comprises e.g. SY₁SGY₂Y₃(SEQ ID NO:443) or an amino acid sequence consisting of the same, wherein Y is₁Is S or G, Y₂Is S or Q, and Y₃Is S or N, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 20, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 21, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 22; or (IV) HCDR1 comprises, e.g., GFTFX₁An amino acid sequence represented by THY (SEQ ID NO:440) or consisting thereof, wherein X₁Is N, S or Q, HCDR2 comprises, for example, ISY₁SGY₂Y₃T (SEQ ID NO:444) or an amino acid sequence consisting thereof, wherein Y ₁Is S or G, Y₂Is S or Q, and Y₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 12, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 23, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 21, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19; (f) (I) HCDR1 contains e.g. GFX₁FSX₂YX₃X₄X₅(SEQ ID NO:445) or a sequence consisting of the amino acid sequence shown in (SEQ ID NO:445), wherein X₁Is S or T, X₂Is S, K or R, X₃Is W or Y, X₄Is I or L, and X₅Is S or N, HCDR2 comprises, e.g., Y₁IY₂QY₃Y₄Y₅EY₆Y₇YVESVKG (SEQ ID NO:446) or a sequence consisting of the same, wherein Y is₁Is S or N, Y₂Is K or H, Y₃Is S, Q or H, Y₄Is G or A, Y₅Is S, H or L, Y₆Is T or K, and Y₇Y, K or R, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises the amino acid sequence shown as SEQ ID NO:237Sequence or consists thereof, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238 and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (II) HCDR1 contains as X₁YX₂X₃X₄(SEQ ID NO:447) or consists of an amino acid sequence as shown in, wherein X₁Is S, K or R, X ₂Is W or Y, X₃Is I or L, and X₄Is S or N, HCDR2 comprises, e.g., Y₁IY₂QY₃Y₄Y₅EY₆Y₇YVESVKG (SEQ ID NO:446) or a sequence consisting of the same, wherein Y is₁Is S or N, Y₂Is K or H, Y₃Is S, Q or H, Y₄Is G or A, Y₅Is S, H or L, Y₆Is T or K, and Y₇Y, K or R, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (III) HCDR1 contains, for example, GFX₁FSX₂Y (SEQ ID NO:448) or an amino acid sequence consisting of the same, wherein X₁Is S or T, and X₂Is S, K or R, HCDR2 comprises e.g. Y₁QY₂Y₃Y₄E (SEQ ID NO:449) or an amino acid sequence consisting of the same, wherein Y₁Is K or H, Y₂Is S, Q or H, Y₃Is G or A, and Y₄S, H or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or (IV) HCDR1 contains, e.g., GFX ₁FSX₂YX₃(SEQ ID NO:450) or consists of an amino acid sequence shown in (SEQ ID NO:450), wherein X₁Is S or T, X₂Is S, K or R, and X₃Is W or Y, HCDR2 comprises, e.g., IY₁QY₂Y₃Y₄EY₅(SEQ ID NO:451) or consists of an amino acid sequence shown in SEQ ID NO,wherein Y is₁Is K or H, Y₂Is S, Q or H, Y₃Is G or A, Y₄Is S, H or L, and Y₅Is T or K, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:232, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:243, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (g) (I) HCDR1 contains e.g. GFX₁FSX₂YX₃X₄X₅(SEQ ID NO:445) or a sequence consisting of the amino acid sequence shown in (SEQ ID NO:445), wherein X₁Is S or T, X₂Is S, K or R, X₃Is W or Y, X₄Is I or L, and X₅Is S or N, HCDR2 comprises e.g. SIHQY₁Y₂Y₃EY₄Y₅YVESVKG (SEQ ID NO:453) or consist of the amino acid sequence shown in, wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, Y₄Is T or K, and Y₅Is K or R, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (II) HCDR1 contains as X ₁YX₂X₃X₄(SEQ ID NO:447) or consists of an amino acid sequence as shown in, wherein X₁Is S, K or R, X₂Is W or Y, X₃Is I or L, and X₄Is S or N, HCDR2 comprises e.g. SIHQY₁Y₂Y₃EY₄Y₅YVESVKG (SEQ ID NO:453) or consist of the amino acid sequence shown in, wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, Y₄Is T or K, and Y₅Is K or R, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (III) HCDR1 contains, for example, GFX₁FSX₂Y (SEQ ID NO:448) or an amino acid sequence consisting of the same, wherein X₁Is S or T, and X₂Is S, K or R, HCDR2 comprises e.g. HQY₁Y₂Y₃E (SEQ ID NO:456) or an amino acid sequence consisting of the same, wherein Y₁Is Q or H, Y₂Is G or A, and Y₃Is H or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or (IV) HCDR1 contains, e.g., GFX ₁FSX₂YX₃(SEQ ID NO:450) or consists of an amino acid sequence shown in (SEQ ID NO:450), wherein X₁Is S or T, X₂Is S, K or R, and X₃Is W or Y, HCDR2 includes, for example, IHQY₁Y₂Y₃EY₄(SEQ ID NO:458) or consists of an amino acid sequence shown in (SEQ ID NO:458) wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, and Y₄Is T or K, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:232, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:243, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (h) (I) HCDR1 comprises, for example, GFTFSX₁YX₂IX₃(SEQ ID NO:452) or consists of an amino acid sequence as shown in (SEQ ID NO:452), wherein X₁Is S or R, X₂Is W or Y, and X₃Is S or N, HCDR2 comprises, e.g., Y₁IY₂QY₃Y₄Y₅EY₆Y₇YVESVKG (SEQ ID NO:446) or a sequence consisting of the same, wherein Y is₁Is S or N, Y₂Is K or H, Y₃Is S, Q or H, Y₄Is G or A, Y₅Is S, H or L, Y₆Is T or K, and Y₇Y, K or R, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, LCDR2 comprises the amino acid sequence shown as SEQ ID NO:238Sequence or consists thereof, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (II) HCDR1 contains as X ₁YX₂IX₃(SEQ ID NO:454) or consists of an amino acid sequence shown in (SEQ ID NO:454), wherein X₁Is S or R, X₂Is W or Y, and X₃Is S or N, HCDR2 comprises, e.g., Y₁IY₂QY₃Y₄Y₅EY₆Y₇YVESVKG (SEQ ID NO:446) or a sequence consisting of the same, wherein Y is₁Is S or N, Y₂Is K or H, Y₃Is S, Q or H, Y₄Is G or A, Y₅Is S, H or L, Y₆Is T or K, and Y₇Y, K or R, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (III) HCDR1 comprises, for example, GFTFSX₁Y (SEQ ID NO:455) or an amino acid sequence consisting thereof, wherein X₁Is S or R, HCDR2 comprises e.g. Y1QY₂Y₃Y₄E (SEQ ID NO:449) or an amino acid sequence consisting of the same, wherein Y₁Is K or H, Y₂Is S, Q or H, Y₃Is G or A, and Y₄S, H or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or (IV) HCDR1 comprises, e.g., GFTFSX ₁YX₂(SEQ ID NO:457) or an amino acid sequence consisting of the amino acid sequence, wherein X₁Is S or R, and X₂Is W or Y, HCDR2 comprises, e.g., IY₁QY₂Y₃Y₄EY₅(SEQ ID NO:451) or consists of an amino acid sequence shown in (SEQ ID NO:451), wherein Y₁Is K or H, Y₂Is S, Q or H, Y₃Is G or A, Y₄Is S, H or L, and Y₅Is T or K, and the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:232Furthermore, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:243, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241 and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; or (I) (I) HCDR1 comprises, e.g., GFTFSX₁YX₂IX₃(SEQ ID NO:452) or consists of an amino acid sequence as shown in (SEQ ID NO:452), wherein X₁Is S or R, X₂Is W or Y, and X₃Is S or N, HCDR2 comprises e.g. SIHQY₁Y₂Y₃EY₄Y₅YVESVKG (SEQ ID NO:453) or consist of the amino acid sequence shown in, wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, Y₄Is T or K, and Y₅Is K or R, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (II) HCDR1 contains as X ₁YX₂IX₃(SEQ ID NO:454) or consists of an amino acid sequence shown in (SEQ ID NO:454), wherein X₁Is S or R, X₂Is W or Y, and X₃Is S or N, HCDR2 comprises e.g. SIHQY₁Y₂Y₃EY₄Y₅YVESVKG (SEQ ID NO:453) or consist of the amino acid sequence shown in, wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, Y₄Is T or K, and Y₅Is K or R, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; (III) HCDR1 comprises, for example, GFTFSX₁Y (SEQ ID NO:455) or an amino acid sequence consisting thereof, wherein X₁Is S or R, HCDR2 comprises e.g. HQY1Y₂Y₃E (SEQ ID NO:456) or an amino acid sequence consisting of the same, wherein Y₁Is Q or H, Y₂Is G or A, and Y₃Is H or L, HCDR3 comprises ammonia as shown in SEQ ID NO:228Amino acid sequence or consists thereof, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241 and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or (IV) HCDR1 comprises, e.g., GFTFSX ₁YX₂(SEQ ID NO:457) or an amino acid sequence consisting of the amino acid sequence, wherein X₁Is S or R, and X₂Is W or Y, HCDR2 includes, for example, IHQY₁Y₂Y₃EY₄(SEQ ID NO:458) or consists of an amino acid sequence shown in (SEQ ID NO:458) wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, and Y₄Is T or K, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:232, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:243, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239.

In some embodiments of the invention, the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3), wherein: (a) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:310, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of GX₁X₂X₃GX₄LGFDH (SEQ ID NO:459) or consists of the amino acid sequence shown in the specification, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:320, LCDR2 comprises, for example, GNSNRPY ₁(SEQ ID NO:460) or consists of an amino acid sequence shown in (SEQ ID NO:460) wherein Y₁Is S or N, and LCDR3 comprises, for example, QSYZ₁Z₂Z₃Z₄Z₅Z₆Z₇V (SEQ ID NO:461) wherein Z is₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:229, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of GX₁X₂X₃GX₄LGFDH (SEQ ID NO:459) or consists of the amino acid sequence shown in the specification, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:320, LCDR2 comprises, for example, GNSNRPY₁(SEQ ID NO:460) or consists of an amino acid sequence shown in (SEQ ID NO:460) wherein Y₁Is S or N, and LCDR3 comprises, for example, QSYZ₁Z₂Z₃Z₄Z₅Z₆Z₇V (SEQ ID NO:461) wherein Z is₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V; (III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:80, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:313, HCDR3 comprises or consists of GX ₁X₂X₃GX₄LGFDH (SEQ ID NO:459) or consists of the amino acid sequence shown in the specification, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:323, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, and LCDR3 comprises or consists of the amino acid sequence shown as YZ₁Z₂Z₃Z₄Z₅Z₆Z₇(SEQ ID NO:462) or an amino acid sequence consisting of the amino acid sequence shown in (SEQ ID NO:462), wherein Z₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V; or (IV) the HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:82, the HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:314, the HCDR3 comprises or consists of ARGX₁X₂X₃GX₄LGFDH(SEQ ID NO:463) The amino acid sequence shown or consisting thereof, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:326, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, and LCDR3 comprises or consists of the amino acid sequence shown as QSYZ₁Z₂Z₃Z₄Z₅Z₆Z₇V (SEQ ID NO:461) wherein Z is₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z ₆Is G, S or F, and Z₇Is P, S or V; (b) (I) HCDR1 comprises e.g. GFTFX₁X₂YAX₃X₄(SEQ ID NO:464) or consists of an amino acid sequence as shown in (SEQ ID NO:464), wherein X₁Is S or G, X₂Is S or T, X₃Is I or M, and X₄Is S or T, HCDR2 comprises e.g. Y₁ISY₂Y₃GY₄Y₅Y₆Y₇YAY₈An amino acid sequence as shown in SVKG (SEQ ID NO:465) or consists thereof, wherein Y₁Is A or S, Y₂Is A, S or G, Y₃Is S or H, Y₄Is G or Y, Y₅Is S or Y, Y₆Is T or A, Y₇Is Y, R or N, and Y₈Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339; (II) HCDR1 contains as X₁YAX₂X₃(SEQ ID NO:466) or consists of an amino acid sequence shown in (SEQ ID NO:466) wherein X₁Is S or T, X₂Is I or M, and X₃Is S or T, HCDR2 comprises e.g. Y₁ISY₂Y₃GY₄Y₅Y₆Y₇YAY₈An amino acid sequence as shown in SVKG (SEQ ID NO:465) or consists thereof, wherein Y₁Is A or S, Y₂Is A, S or G, Y₃Is S or H, Y₄Is G or Y, Y₅Is S or Y, Y₆Is T or A, Y₇Is Y, R or N, and Y₈Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339; (III) HCDR1 comprises e.g. GFTFX ₁X₂Y (SEQ ID NO:467), wherein X is₁Is S or G, and X₂Is S or T, HCDR2 comprises e.g. SY₁Y₂GY₃Y₄(SEQ ID NO:468) or a sequence consisting of the amino acids shown in (SEQ ID NO: Y)₁Is A, S or G, Y₂Is S or H, Y₃Is G or Y, and Y₄Is S or Y, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:340, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 342; or (IV) HCDR1 comprises, e.g., GFTFX₁X₂YA (SEQ ID NO:469) or an amino acid sequence consisting of the same, wherein X₁Is S or G, and X₂Is S or T, HCDR2 comprises, e.g., ISY₁Y₂GY₃Y₄T (SEQ ID NO:470) or an amino acid sequence consisting thereof, wherein Y₁Is S or G, Y₂Is S or H, Y₃Is G or Y, and Y₄Is S or Y, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:332, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:343, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339; or (c) (I) HCDR1 comprises, e.g., GFTFX ₁X₂YAX₃X₄(SEQ ID NO:464) or consists of an amino acid sequence as shown in (SEQ ID NO:464), wherein X₁Is S or G, X₂Is S or T, X₃Is I or M, and X₄Is S or T, HCDR2 comprises e.g. SISY₁Y₂GYYY₃Y₄YAY₅SVKG(SE471) or an amino acid sequence of the formula, wherein Y is₁Is A or S, Y₂Is S or H, Y₃Is T or A, Y₄Is R or N, and Y₅Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339; (II) HCDR1 contains as X₁YAX₂X₃(SEQ ID NO:466) or consists of an amino acid sequence shown in (SEQ ID NO:466) wherein X₁Is S or T, X₂Is I or M, and X₃Is S or T, HCDR2 comprises e.g. SISY₁Y₂GYYY₃Y₄YAY₅An amino acid sequence as shown in SVKG (SEQ ID NO:471) or consists thereof, wherein Y₁Is A or S, Y₂Is S or H, Y₃Is T or A, Y₄Is R or N, and Y₅Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339; (III) HCDR1 comprises e.g. GFTFX ₁X₂Y (SEQ ID NO:467), wherein X is₁Is S or G, and X₂Is S or T, HCDR2 comprises e.g. SY₁Y₂GYY (SEQ ID NO:472) or an amino acid sequence consisting of the same, wherein Y is₁Is A or S, and Y₂Is S or H, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:340, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 342; or (IV) HCDR1 comprises, e.g., GFTFX₁X₂YA (SEQ ID NO:469) or an amino acid sequence consisting of the same, wherein X₁Is S or G, and X₂Is S or T, HCDR2 comprises, e.g., ISY₁Y₂G (SEQ ID NO:473) or an amino acid sequence consisting of the sameWherein Y is₁Is A, S or G, and Y₂Is S or H, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:332, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:343, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339.

In some embodiments of the invention, the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3), wherein: (a) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 28, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 29, 119 and 190, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 30, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 41, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 42, and LCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 43, 126, 134, 145, 172, 178 and 184; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 31, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 29, 119 and 190, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 30, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 41, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 42, and LCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 43, 126, 134, 145, 172, 178 and 184; (III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:32, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID NOs 33, 120 and 191, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:30, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:44, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and LCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID NOs 46, 127, 135, 146, 173, 179 and 185; or (IV) the HCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 34, the HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NO. 35, 121 and 192, the HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 36, the LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 47, the LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 45, and the LCDR3 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NO. 43, 126, 134, 145, 172, 178 and 184; (b) (I) HCDR1 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 4, 112 and 165, HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 5, 100 and 151, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 6, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID No. 17, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID No. 18, and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 19; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 7, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 5, 100 and 151, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 17, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 18, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 19; (III) HCDR1 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 8, 113 and 166, HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 9, 101 and 152, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 6, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID No. 20, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID No. 21 and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 22; or (IV) the HCDR1 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO. 10, 114 and 167, the HCDR2 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO. 11, 102 and 153, the HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 12, the LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 23, the LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 21, and the LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 19; or (c) (I) HCDR1 comprises or consists of an amino acid sequence as set forth in any of SEQ ID NOs: 226, 367 and 378, HCDR2 comprises or consists of an amino acid sequence as set forth in any of SEQ ID NOs: 227, 368 and 379, HCDR3 comprises or consists of an amino acid sequence as set forth in SEQ ID NOs: 228, LCDR1 comprises or consists of an amino acid sequence as set forth in SEQ ID NOs: 237, LCDR2 comprises or consists of an amino acid sequence as set forth in SEQ ID NOs: 238, and LCDR3 comprises or consists of an amino acid sequence as set forth in SEQ ID NOs: 239; (II) HCDR1 comprises or consists of an amino acid sequence as shown in any one of SEQ ID nos. 229, 369 and 380, HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID nos. 227, 368 and 379, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 228, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID No. 237, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID No. 238, and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 239; (III) HCDR1 comprises or consists of the amino acid sequence as shown in any one of SEQ ID nos. 32, 370 and 381, HCDR2 comprises or consists of the amino acid sequence as shown in any one of SEQ ID nos. 230, 371 and 382, HCDR3 comprises or consists of the amino acid sequence as shown in SEQ ID No. 228, LCDR1 comprises or consists of the amino acid sequence as shown in SEQ ID No. 240, LCDR2 comprises or consists of the amino acid sequence as shown in SEQ ID No. 241, and LCDR3 comprises or consists of the amino acid sequence as shown in SEQ ID No. 242; or (IV) HCDR1 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO:34, 372 and 383, HCDR2 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO:231, 373 and 384, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO:232, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO:243, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID NO:241 and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO: 239.

In some embodiments of the invention, the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3), wherein: (a) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:310, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 312 and 348, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:320, LCDR2 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 321 and 354, and LCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 322, 355 and 361; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:229, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 312 and 348, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:320, LCDR2 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 321 and 354, and LCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 322, 355 and 361; (III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:80, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:313, HCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs: 312 and 348, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:323, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, and LCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs: 325, 356 and 362; or (IV) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:82, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:314, HCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID NO:315 and 349, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:326, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, and LCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID NO:322, 355 and 361; or (b) (I) HCDR1 comprises or consists of an amino acid sequence as set forth in any of SEQ ID NO:270 and 407, HCDR2 comprises or consists of an amino acid sequence as set forth in any of SEQ ID NO:271, 389 and 408, HCDR3 comprises or consists of an amino acid sequence as set forth in SEQ ID NO:331, LCDR1 comprises or consists of an amino acid sequence as set forth in SEQ ID NO:337, LCDR2 comprises or consists of an amino acid sequence as set forth in SEQ ID NO:338 and LCDR3 comprises or consists of an amino acid sequence as set forth in SEQ ID NO: 339; (II) HCDR1 comprises or consists of the amino acid sequence shown as any one of SEQ ID NOs 273 and 409, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID NOs 271, 389 and 408, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO 331, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO 337, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO 338, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO 339; (III) HCDR1 comprises or consists of an amino acid sequence as shown in any one of SEQ ID nos. 32 and 410, HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID nos. 274, 390 and 411, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 331, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID No. 340, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID No. 341, and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 342; or (IV) the HCDR1 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO:275 and 412, the HCDR2 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO:276, 391 and 413, the HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO:332, the LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO:343, the LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID NO:341, and the LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO: 339.

In some embodiments of the invention, the antibody or antigen binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from the group consisting of: (a) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 134(LCDR 3); (b) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 126(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:126(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 126(LCDR 3); (c) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 145(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:145(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:146(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 145(LCDR 3); (d) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 172(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 172(LCDR 3); (e) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 178(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:178(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 178(LCDR 3); (f) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 184(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:184(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:185(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 184(LCDR 3); (g) (I) SEQ ID NO 4(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 101(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3); (h) (I) SEQ ID NO:112(HCDR1), SEQ ID NO:100(HCDR2), SEQ ID NO:6(HCDR3), SEQ ID NO:17(LCDR1), SEQ ID NO:18(LCDR2), and SEQ ID NO:19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 113(HCDR1), SEQ ID NO 101(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 114(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3); (i) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 43(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:43(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:46(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 43(LCDR 3); (j) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 126(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:126(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 126(LCDR 3); (k) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 134(LCDR 3); (l) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 145(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:145(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:146(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 145(LCDR 3); (m) (I) SEQ ID NO 4(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3); (n) (I) SEQ ID NO:112(HCDR1), SEQ ID NO:151(HCDR2), SEQ ID NO:6(HCDR3), SEQ ID NO:17(LCDR1), SEQ ID NO:18(LCDR2), and SEQ ID NO:19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 113(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 114(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3); (o) (I) SEQ ID NO:165(HCDR1), SEQ ID NO:151(HCDR2), SEQ ID NO:6(HCDR3), SEQ ID NO:17(LCDR1), SEQ ID NO:18(LCDR2), and SEQ ID NO:19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 166(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO:167(HCDR1), SEQ ID NO:153(HCDR2), SEQ ID NO:12(HCDR3), SEQ ID NO:23(LCDR1), SEQ ID NO:21(LCDR2), and SEQ ID NO:19(LCDR 3); (p) (I) SEQ ID NO:28(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:172(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 172(LCDR 3); (q) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 178(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:178(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 178(LCDR 3); (r) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 184(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:184(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:185(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 184(LCDR 3); (s) (I) SEQ ID NO:28(HCDR1), SEQ ID NO:190(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:190(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:191(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 192(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 134(LCDR 3); (t) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 190(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 172(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:190(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:191(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 192(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 172(LCDR 3); (u) (I) SEQ ID NO 4(HCDR1), SEQ ID NO 5(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 5(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 9(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 11(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3); (v) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 43(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:43(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:46(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2), and SEQ ID NO 43(LCDR 3); (w) (I) SEQ ID NO:367(HCDR1), SEQ ID NO:368(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3); (II) SEQ ID NO:369(HCDR1), SEQ ID NO:368(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3); (III) SEQ ID NO:370(HCDR1), SEQ ID NO:371(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3); or (IV) SEQ ID NO:372(HCDR1), SEQ ID NO:373(HCDR2), SEQ ID NO:232(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:239(LCDR 3); (x) (I) SEQ ID NO:378(HCDR1), SEQ ID NO:379(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3); (II) SEQ ID NO:380(HCDR1), SEQ ID NO:379(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3); (III) SEQ ID NO:381(HCDR1), SEQ ID NO:382(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3); or (IV) SEQ ID NO 383(HCDR1), SEQ ID NO 384(HCDR2), SEQ ID NO 232(HCDR3), SEQ ID NO 243(LCDR1), SEQ ID NO 241(LCDR2), and SEQ ID NO 239(LCDR 3); (y) (I) SEQ ID NO:226(HCDR1), SEQ ID NO:227(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:227(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:230(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3); or (IV) SEQ ID NO:34(HCDR1), SEQ ID NO:231(HCDR2), SEQ ID NO:232(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:239(LCDR 3); (z) (I) SEQ ID NO:270(HCDR1), SEQ ID NO:271(HCDR2), SEQ ID NO:272(HCDR3), SEQ ID NO:282(LCDR1), SEQ ID NO:261(LCDR2), and SEQ ID NO:283(LCDR 3); (II) SEQ ID NO:273(HCDR1), SEQ ID NO:271(HCDR2), SEQ ID NO:272(HCDR3), SEQ ID NO:282(LCDR1), SEQ ID NO:261(LCDR2), and SEQ ID NO:283(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:274(HCDR2), SEQ ID NO:272(HCDR3), SEQ ID NO:284(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:285(LCDR 3); or (IV) SEQ ID NO 275(HCDR1), SEQ ID NO 276(HCDR2), SEQ ID NO 277(HCDR3), SEQ ID NO 286(LCDR1), SEQ ID NO 241(LCDR2), and SEQ ID NO 283(LCDR 3); or (aa) (I) SEQ ID NO:291(HCDR1), SEQ ID NO:292(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:304(LCDR 3); (II) SEQ ID NO:294(HCDR1), SEQ ID NO:292(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:304(LCDR 3); (III) SEQ ID NO:295(HCDR1), SEQ ID NO:296(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:305(LCDR 3); or (IV) SEQ ID NO:297(HCDR1), SEQ ID NO:298(HCDR2), SEQ ID NO:299(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:304(LCDR 3).

In some embodiments of the invention, the antibody or antigen binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from the group consisting of: (a) (I) SEQ ID NO 52(HCDR1), SEQ ID NO 53(HCDR2), SEQ ID NO 54(HCDR3), SEQ ID NO 65(LCDR1), SEQ ID NO 66(LCDR2) and SEQ ID NO 67(LCDR 3); (II) SEQ ID NO:55(HCDR1), SEQ ID NO:53(HCDR2), SEQ ID NO:54(HCDR3), SEQ ID NO:65(LCDR1), SEQ ID NO:66(LCDR2) and SEQ ID NO:67(LCDR 3); (III) SEQ ID NO:56(HCDR1), SEQ ID NO:57(HCDR2), SEQ ID NO:54(HCDR3), SEQ ID NO:68(LCDR1), SEQ ID NO:69(LCDR2), and SEQ ID NO:70(LCDR 3); or (IV) SEQ ID NO 58(HCDR1), SEQ ID NO 59(HCDR2), SEQ ID NO 60(HCDR3), SEQ ID NO 71(LCDR1), SEQ ID NO 69(LCDR2) and SEQ ID NO 67(LCDR 3); (b) (I) SEQ ID NO:76(HCDR1), SEQ ID NO:77(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:89(LCDR1), SEQ ID NO:90(LCDR2) and SEQ ID NO:91(LCDR 3); (II) SEQ ID NO:79(HCDR1), SEQ ID NO:77(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:89(LCDR1), SEQ ID NO:90(LCDR2) and SEQ ID NO:91(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:81(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:92(LCDR1), SEQ ID NO:93(LCDR2), and SEQ ID NO:94(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 83(HCDR2), SEQ ID NO 84(HCDR3), SEQ ID NO 95(LCDR1), SEQ ID NO 93(LCDR2) and SEQ ID NO 91(LCDR 3); (c) (I) SEQ ID NO:310(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2) and SEQ ID NO:361(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:361(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:362(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 314(HCDR2), SEQ ID NO 349(HCDR3), SEQ ID NO 326(LCDR1), SEQ ID NO 324(LCDR2) and SEQ ID NO 361(LCDR 3); (d) (I) SEQ ID NO:270(HCDR1), SEQ ID NO:389(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (II) SEQ ID NO:273(HCDR1), SEQ ID NO:389(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:390(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 275(HCDR1), SEQ ID NO 391(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3); (e) (I) SEQ ID NO:407(HCDR1), SEQ ID NO:408(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (II) SEQ ID NO:409(HCDR1), SEQ ID NO:408(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:410(HCDR1), SEQ ID NO:411(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 412(HCDR1), SEQ ID NO 413(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3); (f) (I) SEQ ID NO:310(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:321(LCDR2) and SEQ ID NO:322(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:321(LCDR2), and SEQ ID NO:322(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:325(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 314(HCDR2), SEQ ID NO 315(HCDR3), SEQ ID NO 326(LCDR1), SEQ ID NO 324(LCDR2) and SEQ ID NO 322(LCDR 3); (g) (I) SEQ ID NO 270(HCDR1), SEQ ID NO 271(HCDR2), SEQ ID NO 331(HCDR3), SEQ ID NO 337(LCDR1), SEQ ID NO 338(LCDR2) and SEQ ID NO 339(LCDR 3); (II) SEQ ID NO:273(HCDR1), SEQ ID NO:271(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:274(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 275(HCDR1), SEQ ID NO 276(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3); or (h) (I) SEQ ID NO 310(HCDR1), SEQ ID NO 311(HCDR2), SEQ ID NO 348(HCDR3), SEQ ID NO 320(LCDR1), SEQ ID NO 354(LCDR2), and SEQ ID NO 355(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:355(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2) and SEQ ID NO:356(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 314(HCDR2), SEQ ID NO 349(HCDR3), SEQ ID NO 326(LCDR1), SEQ ID NO 324(LCDR2) and SEQ ID NO 355(LCDR 3).

In some embodiments of the invention, the antibody or antigen-binding fragment comprises: (a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 136; (b) 122, and a light chain variable region comprising the amino acid sequence of SEQ ID No. 136; (c) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 128; (d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 128; (e) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 147; (f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 147; (g) (ii) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO:201 and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 174; (h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 174; (i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 180; (j) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 180; (k) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 186; (l) 122, and a light chain variable region comprising the amino acid sequence of SEQ ID NO 186; (m) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:103 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24; (n) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:115 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24; (o) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 48; (p) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 128; (q) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 136; (r) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 147; (s) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:154 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24; (t) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:161 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24; (u) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:168, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24; (v) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 174; (w) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 180; (x) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO 37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 186; (y) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:193 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 136; (z) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:193, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 174; (aa) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:13 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24; (bb) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 48; (cc) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:374, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 244; (dd) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:385 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 244; (ee) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO:233, and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 244; (ff) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO:278 and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 287; or (gg) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:300 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 306.

In some embodiments of the invention, the antibody or antigen-binding fragment comprises: (a) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO 61 and the light chain variable region comprising the amino acid sequence of SEQ ID NO 72; (b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 85 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 96; (c) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:350 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 363; (d) 392 and 344; (e) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 414 and the light chain variable region comprising the amino acid sequence of SEQ ID NO. 344; (f) 316, and a light chain variable region comprising the amino acid sequence of SEQ ID No. 327; (g) (ii) a heavy chain variable region comprising the amino acid sequence of SEQ ID No. 333 and a light chain variable region comprising the amino acid sequence of SEQ ID No. 344; or (h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:350 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 357.

In some embodiments of the invention, the antibody or antigen-binding fragment comprises: (a) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 138; (b) a heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 138; (c) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 130; (d) a heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 130; (e) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 149; (f) a heavy chain comprising the amino acid sequence of SEQ ID NO:208 and a light chain comprising the amino acid sequence of SEQ ID NO: 149; (g) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 176; (h) a heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 176; (i) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 182; (j) a heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 182; (k) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 188; (l) A heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 188; (m) a heavy chain comprising the amino acid sequence of SEQ ID NO:105 and a light chain comprising the amino acid sequence of SEQ ID NO: 26; (n) a heavy chain comprising the amino acid sequence of SEQ ID NO:108 and a light chain comprising the amino acid sequence of SEQ ID NO: 26; (o) a heavy chain comprising the amino acid sequence of SEQ ID NO:117 and a light chain comprising the amino acid sequence of SEQ ID NO: 26; (p) a heavy chain comprising the amino acid sequence of SEQ ID NO:124 and a light chain comprising the amino acid sequence of SEQ ID NO: 50; (q) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 130; (r) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 138; (s) a heavy chain comprising the amino acid sequence of SEQ ID NO:141 and a light chain comprising the amino acid sequence of SEQ ID NO: 138; (t) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 149; (u) a heavy chain comprising the amino acid sequence of SEQ ID NO:156 and a light chain comprising the amino acid sequence of SEQ ID NO: 26; (v) a heavy chain comprising the amino acid sequence of SEQ ID NO. 159 and a light chain comprising the amino acid sequence of SEQ ID NO. 26; (w) a heavy chain comprising the amino acid sequence of SEQ ID NO:163 and a light chain comprising the amino acid sequence of SEQ ID NO: 26; (x) A heavy chain comprising the amino acid sequence of SEQ ID NO:170 and a light chain comprising the amino acid sequence of SEQ ID NO: 26; (y) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 176; (z) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 182; (aa) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 188; (bb) a heavy chain comprising the amino acid sequence of SEQ ID NO:195, and a light chain comprising the amino acid sequence of SEQ ID NO: 138; (cc) a heavy chain comprising the amino acid sequence of SEQ ID NO:195, and a light chain comprising the amino acid sequence of SEQ ID NO: 176; (dd) a heavy chain comprising the amino acid sequence of SEQ ID NO:15 and a light chain comprising the amino acid sequence of SEQ ID NO: 26; (ee) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 50; (ff) a heavy chain comprising the amino acid sequence of SEQ ID NO:376 and a light chain comprising the amino acid sequence of SEQ ID NO: 246; (gg) a heavy chain comprising the amino acid sequence of SEQ ID NO:387 and a light chain comprising the amino acid sequence of SEQ ID NO: 246; (hh) a heavy chain comprising the amino acid sequence of SEQ ID NO:235 and a light chain comprising the amino acid sequence of SEQ ID NO: 246; (ii) a heavy chain comprising the amino acid sequence of SEQ ID NO 280 and a light chain comprising the amino acid sequence of SEQ ID NO 289; or (jj) a heavy chain comprising the amino acid sequence of SEQ ID NO:302 and a light chain comprising the amino acid sequence of SEQ ID NO: 308.

In some embodiments of the invention, the antibody or antigen-binding fragment comprises: (a) a heavy chain comprising the amino acid sequence of SEQ ID NO 63 and a light chain comprising the amino acid sequence of SEQ ID NO 74; (b) a heavy chain comprising the amino acid sequence of SEQ ID NO:87 and a light chain comprising the amino acid sequence of SEQ ID NO: 98; (c) a heavy chain comprising the amino acid sequence of SEQ ID NO 352 and a light chain comprising the amino acid sequence of SEQ ID NO 365; (d) a heavy chain comprising the amino acid sequence of SEQ ID NO. 394 and a light chain comprising the amino acid sequence of SEQ ID NO. 346; (e) a heavy chain comprising the amino acid sequence of SEQ ID NO 416, and a light chain comprising the amino acid sequence of SEQ ID NO 346; (f) a heavy chain comprising the amino acid sequence of SEQ ID NO:318, and a light chain comprising the amino acid sequence of SEQ ID NO: 329; (g) a heavy chain comprising the amino acid sequence of SEQ ID NO 335 and a light chain comprising the amino acid sequence of SEQ ID NO 346; or (h) a heavy chain comprising the amino acid sequence of SEQ ID NO:352 and a light chain comprising the amino acid sequence of SEQ ID NO: 359.

In some embodiments of the invention, the antibody or antigen-binding fragment is an antigen-binding fragment selected from the group consisting of: fab, Fab ', F (ab') ₂Fv, single domain antibodies (dAbs), and single chain variable fragments (scFv). In some embodiments of the invention, the antibody or antigen-binding fragment is an antigen-binding fragment selected from the group consisting of: fab, Fab', Fv, single junctionDomain antibodies (dabs) and single chain variable fragments (scFv).

In some embodiments of the invention, the antibody or antigen-binding fragment is monoclonal. In some embodiments of the invention, the antibody or antigen-binding fragment is fully human. In some embodiments of the invention, the antibody or antigen-binding fragment is an IgG antibody. In some embodiments of the invention, the antibody or antigen-binding fragment is an IgG1 antibody. In some embodiments of the invention, the antibody or antigen-binding fragment is an IgG1 antibody with a kappa light chain. In some embodiments of the invention, the antibody or antigen-binding fragment is a fully human antibody of the IgG1 isotype, with a kappa light chain.

In some embodiments of the invention, the antibody or antigen binding fragment additionally has a mutation in the Fc region according to the EU index of Kabat (Kabat), wherein the mutation comprises at least D265A and P329A.

In some embodiments of the invention, the antibody or antigen-binding fragment additionally has a mutation in the Fc region according to the EU index of kabat, wherein the mutation comprises at least L234A and L235A.

In some embodiments of the invention, the antibody or antigen binding fragment is therapeutic.

In some embodiments of the invention, the antibody or antigen-binding fragment binds to the same epitope on human NPR1 as any antibody or antigen-binding fragment or group (e.g., XX16) defined herein. In some embodiments of the invention, the antibody or antigen-binding fragment competes with any antibody or antigen-binding fragment or group (e.g., XX16) defined herein for binding to human NPR 1.

In one aspect of the invention, provided herein is one or more isolated nucleic acids encoding the amino acid sequence of any of the antibodies or antigen-binding fragments or groups defined herein. In one aspect of the invention, provided herein are vectors comprising the one or more isolated nucleic acids. In one aspect of the invention, provided herein are host cells comprising the one or more isolated nucleic acids or vectors.

In one aspect of the invention, provided herein is a method of producing any of the antibodies or antigen-binding fragments described herein, comprising culturing a host cell described herein under conditions suitable for production of the antibody or antigen-binding fragment. In some embodiments of the invention, the method additionally comprises purifying the antibody or antigen-binding fragment.

In one aspect of the invention, provided herein are pharmaceutical compositions comprising a purified antibody or antigen-binding fragment produced by the methods described herein and a pharmaceutically acceptable carrier.

In one aspect of the invention, provided herein is a pharmaceutical composition comprising any of the antibodies or antigen-binding fragments described herein and a pharmaceutically acceptable carrier.

In one aspect of the present invention, provided herein is a pharmaceutical composition comprising: a) a device that binds to natriuretic peptide receptor 1(NPR1) and activates NPR1 in the absence of ANP; and b) a pharmaceutically acceptable excipient. In some embodiments of the invention, the means for binding and activating is ANP non-competitive. In some embodiments of the invention, the means for binding and activating is ANP competitive. In some embodiments of the invention, the means for binding and activating is additionally capable of stabilizing the ANP-NPR1 complex. In some embodiments of the invention, the composition further comprises an additional therapeutic agent.

In some embodiments of the invention, the additional therapeutic agent is selected from ACE (angiotensin converting enzyme) inhibitors, Angiotensin Receptor Blockers (ARBs), enkephalinase inhibitors, beta blockers, diuretics, calcium channel blockers, cardiac glycosides, sodium-glucose cotransporter 2 inhibitors (SGLT2i), and combinations thereof. In some embodiments of the invention, the additional therapeutic agent is selected from enalapril, benazepril, captopril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, trandolapril, valsartan, azilsartan, candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, sabotaltraz, bisoprolol, carvedilol, propranolol, metoprolol tartrate, metoprolol succinate, thiazide diuretics, loop diuretics, potassium sparing diuretics, amlodipine, clevidipine, diltiazem, felodipine, isradipine, nicardipine, nifedipine, nisoldipine, verapamil, digitoside, canagliflozin, dapagliflozin, eprogliflozin, and combinations thereof. In some embodiments of the invention, the additional therapeutic agent is selected from the group consisting of chlorothiazide, chlorthalidone, hydrochlorothiazide, indapamide, metolazone, bumetanide, ethacrynic acid, furosemide, torasemide, amiloride, eplerenone, spironolactone, triamterene, digoxin, and combinations thereof. In some embodiments of the invention, the additional therapeutic agent is an angiotensin receptor-enkephalinase inhibitor (ARNi).

In some embodiments of the invention, the additional therapeutic agent is selected from the group consisting of corticosteroids, leukotriene modifiers, bronchodilators, and combinations thereof. In some embodiments of the invention, the additional therapeutic agent is selected from the group consisting of fluticasone, budesonide, mometasone, beclomethasone, ciclesonide, fluticasone furoate, prednisone, methylprednisolone, montelukast, zafirlukast, zileuton, long-acting beta agonists, short-acting beta agonists, theophylline, and ipratropium, and combinations thereof. In some embodiments of the invention, the additional therapeutic agent is selected from salmeterol, formoterol, salbutamol and levalbuterol and combinations thereof.

In some embodiments of the invention, the additional therapeutic agent is selected from the group consisting of a beta-adrenoreceptor antagonist, a carbonic anhydrase inhibitor, an alpha 2-adrenoreceptor agonist, a parasympathomimetic, a prostaglandin analog, a rho kinase inhibitor, and combinations thereof. In some embodiments of the invention, the additional therapeutic agent is selected from the group consisting of timolol, levobunolol, metiprolol, carteolol, betaxolol, acetazolamide, dorzolamide, brinzolamide, methazolamide, brimonidine, alaclonidine, a cholinomide, latanoprost (latanoprost), latanoprost (latanoproste bund), travoprost, bimatoprost, tafluprost, nervosudil (netarsudil), and lapacholide, and combinations thereof.

In one aspect of the invention, provided herein is a method of treating a disorder or disease associated with natriuretic peptide receptor activity in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any of the antibodies or antigen binding fragments described herein or a pharmaceutical composition or combination as described herein.

In one aspect of the invention, provided herein is a method of treating a cardiovascular disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the antibodies or antigen-binding fragments thereof described herein or any one of the pharmaceutical compositions or combinations described herein.

In some embodiments of the invention, the cardiovascular disorder is selected from: hypertension, peripheral vascular disease, heart failure, Coronary Artery Disease (CAD), Ischemic Heart Disease (IHD), mitral stenosis and regurgitation, angina, hypertrophic cardiomyopathy, diabetic cardiomyopathy, supraventricular and ventricular arrhythmias, arrhythmia, Atrial Fibrillation (AF), new atrial fibrillation, recurrent atrial fibrillation, cardiac fibrosis, atrial flutter, detrimental vascular remodeling, plaque stabilization and Myocardial Infarction (MI).

In one aspect of the invention, provided herein is a method of treating heart failure, Hypertrophic Cardiomyopathy (HCM), hypertension, preeclampsia, asthma, glaucoma, and/or cytokine release syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the antibodies or antigen-binding fragments thereof described herein or any one of the pharmaceutical compositions or combinations described herein.

In some embodiments of the invention, the subject has heart failure, wherein the heart failure is selected from heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), heart failure after acute myocardial infarction, or acute compensated heart failure. In some embodiments of the invention, the subject has hypertrophic cardiomyopathy, wherein the hypertrophic cardiomyopathy is ventricular hypertrophy. In some embodiments of the invention, the subject has hypertension, wherein the hypertension is selected from refractory hypertension, hypertensive heart disease, pulmonary hypertension, pulmonary arterial hypertension, isolated systolic hypertension, refractory hypertension, and pulmonary arterial hypertension. In some embodiments of the invention, the subject has hypertension, wherein the hypertension is selected from refractory hypertension or hypertensive heart disease.

In one aspect of the invention, provided herein is a method of treating a renal disorder in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of any one of the antibodies or antigen-binding fragments thereof described herein or any one of the pharmaceutical compositions or combinations described herein. In some embodiments of the invention, the renal disorder is selected from: diabetic renal insufficiency, non-diabetic renal insufficiency, renal failure, diabetic nephropathy, non-diabetic nephropathy, acute kidney injury, contrast-induced nephropathy, nephrotic syndrome, glomerulonephritis, scleroderma, glomerulosclerosis, primary nephrotic proteinuria, renovascular hypertension, diabetic retinopathy and end-stage renal disease (ESRD), endothelial dysfunction, diastolic dysfunction, renal fibrosis and Polycystic Kidney Disease (PKD).

In one aspect of the invention, provided herein is the use of any one of the antibodies or antigen-binding fragments thereof described herein or any one of the pharmaceutical compositions or combinations described herein for the manufacture of a medicament for the treatment of a disorder or disease associated with natriuretic peptide receptor activity in a subject in need of such treatment.

In one aspect of the invention, provided herein is the use of any one of the antibodies or antigen-binding fragments thereof described herein or any one of the pharmaceutical compositions or combinations described herein for the manufacture of a medicament for the treatment of a cardiovascular disorder in a subject in need of such treatment.

In one aspect of the invention, provided herein is the use of any one of the antibodies or antigen-binding fragments thereof described herein or any one of the pharmaceutical compositions or combinations described herein for the manufacture of a medicament for the treatment of heart failure, Hypertrophic Cardiomyopathy (HCM), hypertension, preeclampsia, asthma, glaucoma, and/or cytokine release syndrome in a subject in need of such treatment.

In some embodiments of the invention, the subject has heart failure, and the heart failure is selected from heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), heart failure after acute myocardial infarction, or acute compensated heart failure. In some embodiments of the invention, the subject has hypertrophic cardiomyopathy, wherein the hypertrophic cardiomyopathy is ventricular hypertrophy. In some embodiments of the invention, the subject has hypertension, wherein the hypertension is selected from refractory hypertension, hypertensive heart disease, pulmonary hypertension, pulmonary arterial hypertension, isolated systolic hypertension, refractory hypertension, and pulmonary arterial hypertension. In some embodiments of the invention, the subject has hypertension, wherein the hypertension is selected from refractory hypertension or hypertensive heart disease.

In one aspect of the invention, provided herein is the use of any one of the antibodies or antigen-binding fragments thereof described herein or any one of the pharmaceutical compositions or combinations described herein for the manufacture of a medicament for the treatment of a renal disorder in a subject in need of such treatment. In some embodiments of the invention, the renal disorder is selected from: diabetic renal insufficiency, non-diabetic renal insufficiency, renal failure, diabetic nephropathy, non-diabetic nephropathy, acute kidney injury, contrast-induced nephropathy, nephrotic syndrome, glomerulonephritis, scleroderma, glomerulosclerosis, primary nephrotic proteinuria, renovascular hypertension, diabetic retinopathy and end-stage renal disease (ESRD), endothelial dysfunction, diastolic dysfunction, renal fibrosis and Polycystic Kidney Disease (PKD).

In one aspect of the invention, provided herein is any one of the antibodies or antigen-binding fragments thereof described herein or the pharmaceutical compositions or combinations described herein for use in treating a disorder or disease associated with natriuretic peptide receptor activity in a subject in need of such treatment.

In one aspect of the invention, provided herein is any one of the antibodies or antigen-binding fragments thereof described herein or the pharmaceutical compositions or combinations described herein for use in treating a cardiovascular disorder in a subject in need of such treatment.

In one aspect of the invention, provided herein is any one of the antibodies or antigen-binding fragments thereof described herein or the pharmaceutical compositions or combinations described herein for use in the treatment of heart failure, Hypertrophic Cardiomyopathy (HCM), hypertension, preeclampsia, asthma, glaucoma, and/or cytokine release syndrome in a subject in need of such treatment.

In one aspect of the invention, provided herein is any one of the antibodies or antigen-binding fragments thereof described herein or the pharmaceutical compositions or combinations described herein for use in treating a renal disorder in a subject in need of such treatment. In some embodiments of the invention, the renal disorder is selected from: diabetic renal insufficiency, non-diabetic renal insufficiency, renal failure, diabetic nephropathy, non-diabetic nephropathy, acute kidney injury, contrast-induced nephropathy, nephrotic syndrome, glomerulonephritis, scleroderma, glomerulosclerosis, primary nephrotic proteinuria, renovascular hypertension, diabetic retinopathy and end-stage renal disease (ESRD), endothelial dysfunction, diastolic dysfunction, renal fibrosis and Polycystic Kidney Disease (PKD).

In one aspect of the invention, provided herein is a method of treating a disorder or disease associated with natriuretic peptide receptor activity in a subject in need thereof, the method comprising administering a pharmaceutical composition comprising: a device that binds to natriuretic peptide receptor 1(NPR1) and activates NPR1 in the absence of ANP; and a pharmaceutically acceptable excipient.

In one aspect of the present invention, provided herein is a method of treating a cardiovascular disorder in a subject in need thereof, the method comprising administering a pharmaceutical composition comprising: a device that binds to natriuretic peptide receptor 1(NPR1) and activates NPR1 in the absence of ANP; and a pharmaceutically acceptable excipient. In some embodiments of the invention, the cardiovascular disorder is selected from: hypertension, peripheral vascular disease, heart failure, Coronary Artery Disease (CAD), Ischemic Heart Disease (IHD), mitral stenosis and regurgitation, angina, hypertrophic cardiomyopathy, diabetic cardiomyopathy, supraventricular and ventricular arrhythmias, arrhythmia, Atrial Fibrillation (AF), new atrial fibrillation, recurrent atrial fibrillation, cardiac fibrosis, atrial flutter, detrimental vascular remodeling, plaque stabilization and Myocardial Infarction (MI).

In one aspect of the present invention, provided herein is a method of treating heart failure, Hypertrophic Cardiomyopathy (HCM), hypertension, preeclampsia, asthma, glaucoma, and/or cytokine release syndrome in a subject in need thereof, the method comprising administering a pharmaceutical composition comprising: a device that binds to natriuretic peptide receptor 1(NPR1) and activates NPR1 in the absence of ANP; and a pharmaceutically acceptable excipient. In some embodiments of the invention, the subject has heart failure, wherein the heart failure is selected from heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), heart failure after acute myocardial infarction, or acute compensated heart failure. In some embodiments of the invention, the subject has hypertrophic cardiomyopathy, wherein the hypertrophic cardiomyopathy is ventricular hypertrophy. In some embodiments of the invention, the subject has hypertension, wherein the hypertension is selected from refractory hypertension, hypertensive heart disease, pulmonary hypertension, pulmonary arterial hypertension, isolated systolic hypertension, refractory hypertension, and pulmonary arterial hypertension. In some embodiments of the invention, the subject has hypertension, wherein the hypertension is selected from refractory hypertension or hypertensive heart disease.

In one aspect of the present invention, provided herein is a method of treating a renal disorder in a subject in need thereof, the method comprising administering a pharmaceutical composition comprising: a device that binds to natriuretic peptide receptor 1(NPR1) and activates NPR1 in the absence of ANP; and a pharmaceutically acceptable excipient. In some embodiments of the invention, the renal disorder is selected from: diabetic renal insufficiency, non-diabetic renal insufficiency, renal failure, diabetic nephropathy, non-diabetic nephropathy, acute kidney injury, contrast-induced nephropathy, nephrotic syndrome, glomerulonephritis, scleroderma, glomerulosclerosis, primary nephrotic proteinuria, renovascular hypertension, diabetic retinopathy and end-stage renal disease (ESRD), endothelial dysfunction, diastolic dysfunction, renal fibrosis and Polycystic Kidney Disease (PKD). In some embodiments of the invention, the means for binding and activating is ANP non-competitive. In some embodiments of the invention, the means for binding and activating is ANP competitive. In some embodiments of the invention, the means for binding and activating is additionally capable of stabilizing the ANP-NPR1 complex.

Drawings

Figure 1 is a set of graphs (ELISA analysis) showing the results of binding of antibody candidates WW01, WW02, WW03, WW04 and WW06 to the following antigens in the absence and presence of a 250-fold molar excess of ANP: human NPR1, constitutively active human NPR1 mutant (W74R), rat NPR1 and human NPR3 (reverse target).

FIG. 2 is a set of graphs showing the results of flow cytometric analysis of antibody candidates WW01, WW02, WW03, WW04 and WW06 for binding to CHO-K1 cells expressing human NPR1 in the absence and presence of saturating concentrations of ANP and binding on parental CHO-K1 cells.

Figure 3 is a graphical representation of a Fluorescence Resonance Energy Transfer (FRET) based assay in which NPR 1-specific antibodies compete with ANP for binding to NPR 1. In this FRET-based assay, Eu-labeled streptavidin (for measurement of IgG) or Eu-labeled anti-hFc antibody (for measurement of FabCys) was used as the energy donor, while Cy 5-labeled ANP was used as the acceptor.

Fig. 4 is a set of graphs showing the results of ANP competition analysis on candidates WW01, WW02, WW03, WW04 and WW06 using the FRET-based assay shown in fig. 3.

FIG. 5 is a set of graphs depicting the results of functional activity assays of candidates WW01, WW02, WW03, WW04 and WW06 in a cellular cGMP production assay using CHO-K1 cells expressing human NPR 1. The results represent cellular production of cGMP [ nM ] in the absence or presence of 0.075nM ANP.

FIG. 6 is a set of graphs depicting the results of an assay for the functional activity of the candidate WW06 in the form of IgG or FabCys in a cellular cGMP production assay using CHO-K1 cells expressing human NPR 1. The results represent cellular production of cGMP [ nM ] in the absence or presence of 0.075nM ANP.

FIG. 7 shows that 82 parental antibody based HCDR2 or LCDR3 are only improved

SET screening (hNPR1 affinity) results for derivatives.

FIG. 8 shows that 82 parental antibody based HCDR2 or LCDR3 are only improved

SET screening (hNPR1-ANP complex affinity) results of the derivatives.

Figure 9 is a set of graphs (ELISA analysis) showing the results of binding of antibody candidates XX01-XX08, XX10 and XX12 to the following antigens in the absence or presence of a 250-fold molar excess of ANP: human NPR1, constitutively active human NPR1 mutant (W74R), rat NPR1 and human NPR3 (reverse target).

FIG. 10 is a set of graphs showing the results of flow cytometric analysis of antibody candidates XX01-XX08, XX10, and XX12 for binding to CHO-K1 cells expressing human NPR1 in the absence or presence of saturating concentrations of ANP and binding on parental CHO-K1 cells.

FIG. 11 is a set of graphs showing the results of ANP competition analysis for candidates XX01-XX07, XX10 and XX12 using the FRET-based assay shown in FIG. 3.

FIG. 12 is a set of graphs depicting the results of an assay of the functional activity of candidates XX01-XX08, XX10 and XX12 in a cellular cGMP production assay using CHO-K1 cells expressing human NPR 1. The results represent cellular production of cGMP [ nM ] in the absence or presence of 0.075nM ANP.

FIG. 13 is a set of graphs depicting the results of functional activity assays of candidates XX06 and XX16 and the natural ligand ANP in a cellular cGMP production assay using CHO-K1 cells expressing human NPR 1. The results represent cellular production of cGMP [ nM ] in the absence or presence of 0.075nM ANP.

FIG. 14 is a graph depicting the results of an assay of the functional activity of candidate XX16 and the natural ligand ANP in a cellular cGMP production assay using CHO-K1 cells expressing human NPR 1. The results represent cellular production of cGMP [ nM ] as a function of ANP or XX 16.

Fig. 15 is a schematic representation of the crystal structure of Fab06 (the Fab form of the WW06 antibody) complexed to the ectodomain of hNPR 1.

Figure 16 is a graphical representation of the conformation of the extracellular domain of hNPR1, as if complexed with Fab 06. Fab06 was removed from the graph to more clearly reveal the conformation of hNPR1 induced by Fab binding. W74R is represented as space filling.

Figure 17 is a graphic representation of the crystal structure of the hNPR1 ectodomain complexed with ANP (left) and the crystal structure of Fab16 (XX 16 antibody in Fab form) complexed with the hNPR1 ectodomain (right), where W74 is represented as space filling.

FIG. 18 is a set of graphs depicting the results of an assay of the functional activity of candidate WW06 in a cellular cGMP production assay when tested on CHO-K1 cells expressing hNPR 1W 74R/C232T (constitutively active mutant; Panel A) when compared to WT hNPR1 (Panel B).

Figure 19 is a set of graphs (ELISA assay) showing the results of binding of antibody candidates YY01-YY07 to the following antigens in the absence or presence of 250-fold molar excess of ANP: human NPR1, constitutively active human NPR1 mutant (W74R), rat NPR1 and human NPR3 (reverse target).

FIG. 20 is a set of graphs showing the results of flow cytometric analysis of antibody candidates YY01-YY07 for binding to CHO-K1 cells expressing human NPR1 in the absence or presence of saturating concentrations of ANP and binding on parental CHO K1 cells.

FIG. 21 is a set of graphs showing the results of ANP competition analysis for candidates YY01-YY07 using the FRET-based assay shown in FIG. 3.

FIG. 22 is a set of graphs depicting the results of an assay for the functional activity of candidate YY01-YY07 in a cellular cGMP production assay using CHO-K1 cells expressing human NPR 1. The results represent cellular production of cGMP [ nM ] in the absence or presence of 0.075nM ANP.

FIG. 23 is a set of graphs depicting the results of an assay of the functional activity of candidates YY05 and YY07 in the form of IgG or FabCys in a cellular cGMP production assay using CHO-K1 cells expressing human NPR 1. The results represent cellular production of cGMP [ nM ] in the absence or presence of 0.075nM ANP.

FIG. 24 shows that 112 parental antibody-based HCDR1/2 or LCDR3 only improved

SET screening (hNPR1 affinity) results for derivatives.

FIG. 25 shows that 112 parental antibody based HCDR1/2 or LCDR3 only improved

SET screening (hNPR1-ANP complex affinity) results of the derivatives.

FIG. 26 is a set of graphs showing plasma cGMP (nM) concentration as a function of time in hNPR1 Tg mice injected intravenously with 1mg/kg, 3mg/kg, or 10mg/kg WW06 Fab.

FIG. 27 is a graph showing the change in antibody concentration over time in hNPR1 Tg mice injected intravenously with 1mg/kg, 3mg/kg, or 10mg/kg WW06 Fab.

FIG. 28 is a set of graphs showing the results on heart weight/body weight ratio (left) and plasma NT-proBNP levels (pg/mL; right) in wild type or ANP knockout (ANP KO) mice administered vehicle subcutaneously 0.3mg/kg XX16 or 3mg/kg XX16 once every two weeks. Measurements were taken two weeks after the second application.

FIG. 29 is a set of graphs showing the results with respect to blood pressure (mean arterial pressure; left) and uroflow rate (right) in hypertensive rats (stroke prone spontaneous hypertensive rats, SHRsp) with a single intravenous administration of vehicle, 0.3mg/kg XX16 or 1mg/kg XX 16. Measurements were taken three hours after intravenous administration.

FIG. 30 is a set of graphs showing the results of a single subcutaneous administration of 0.1, 0.3, 1, 3, 10 or 30mg/kg XX16 in telemetrically implanted normal rats, with respect to mean arterial pressure (MAP; upper graph) and plasma cGMP (lower graph) as a function of time.

FIG. 31 is a set of graphs depicting the results of functional activity assays of candidate WW03 and a control antibody (antibody 5591-IgG from PCT application No. WO 2010/065293A 1) in a cellular cGMP production assay using CHO-K1 cells expressing human NPR1 or rat cells expressing rat NPR 1. The results represent cellular production of cGMP [ nM ] in the absence or presence of 0.075nM ANP (human or rat).

General definition

In order that the disclosure may be more readily understood, certain terms are first defined. Additional definitions are set forth throughout the detailed description as appropriate.

Throughout the description and claims of this specification, the words "comprise" and "contain" and variations of the words, for example "comprising" and "comprises", mean "including but not limited to", and do not exclude other components, integers or steps. Furthermore, the singular encompasses the plural unless the context otherwise requires: in particular, where the indefinite article is used, the specification is to be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

As used herein, "NPR 1" and "NPR 1 protein" refer to natriuretic peptide receptor 1. The protein is also known as the atrial natriuretic peptide receptor type A (ANP-A, ANPR-A or NPR-A) and guanylate cyclase A (GC-A). In some embodiments, the mentioned NPR1 is human NPR 1. In some embodiments, human NPR1 has UniProt accession number P16066 or GenBank accession number EAW53284.1(SEQ ID NO: 1). In some embodiments, the mentioned NPR1 is mouse (mus musculus) NPR 1. In some embodiments, the mouse NPR1 has NCBI reference sequence number NP-032753.5 (SEQ ID NO: 2). In some embodiments, the mentioned NPR1 is rat (rattus norvegicus) NPR 1. In some embodiments, rat NPR1 has NCBI reference sequence number NP-036745.1 (SEQ ID NO: 3). Exemplary NPR1 proteins are shown in table 1. In the context of the discussion of constitutive activity or W74R mutants, the mutant refers to Trp at amino acid 74 of the mature human NPR1 protein, which may also be referred to as Trp at amino acid 106 of the hNPR1 protein as shown in SEQ ID NO: 1.

TABLE 1 NPR1 protein sequences

In various embodiments, the anti-NPR 1 antibodies and antigen binding fragments disclosed herein are capable of binding to NPR1 and activating NPR1 in the absence of ANP. With this activity, the disclosed anti-NPR 1 antibodies and antigen-binding fragments can be used to treat undesirable conditions, diseases and disorders, including cardiovascular disorders (e.g., hypertension, peripheral vascular disease, heart failure (including, but not limited to, heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), post-acute myocardial infarction heart failure or acute compensatory heart failure), coronary heart disease (CAD), Ischemic Heart Disease (IHD), mitral stenosis and regurgitation, angina pectoris, hypertrophic cardiomyopathy (e.g., ventricular hypertrophy), diabetic cardiomyopathy, supraventricular and ventricular arrhythmias, arrhythmia, Atrial Fibrillation (AF), new atrial fibrillation, recurrent atrial fibrillation, cardiac fibrosis, atrial flutter, impaired vascular remodeling, plaque stabilization or Myocardial Infarction (MI)), hypertension (refractory hypertension, hypertensive heart disease), Pulmonary hypertension, pulmonary arterial hypertension, isolated systolic hypertension, refractory hypertension or pulmonary arterial hypertension), preeclampsia, asthma, glaucoma, cytokine release syndrome and/or renal disorders (e.g., diabetic renal insufficiency, non-diabetic renal insufficiency, renal failure, diabetic nephropathy, non-diabetic nephropathy, acute kidney injury, contrast-induced nephropathy, nephrotic syndrome, glomerulonephritis, scleroderma, glomerulosclerosis, idiopathic nephrotic proteinuria, renovascular hypertension, diabetic retinopathy and End Stage Renal Disease (ESRD), endothelial dysfunction, diastolic dysfunction, renal fibrosis and Polycystic Kidney Disease (PKD)).

As used herein, the term "antibody" refers to a whole antibody or an antigen-binding fragment thereof. Whole antibodies are glycoproteins comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region consists of three domains, CH1, CH2, and CH 3. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists ofOne domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed Complementarity Determining Regions (CDRs), between which more conserved regions, termed Framework Regions (FRs), are interspersed. Each VH and VL is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR 4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant region of an antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component of the classical complement system (Clq). The term "antibody" includes, but is not limited to, monoclonal antibodies, human antibodies, humanized antibodies, camelized (camelized) antibodies, and chimeric antibodies. The antibody can be of any isotype/class (e.g., IgG, IgE, IgM, IgD, IgA, and IgY) or subclass (e.g., IgG) ₁、IgG₂、IgG₃、IgG₄、IgA₁And IgA₂)。

The term "antigen-binding fragment" refers to a fragment of an intact antibody that retains the ability to specifically bind a given antigen (e.g., NPR1) and/or provide the function of the intact antibody. Such fragments include Fab fragments, Fab' fragments, which are monovalent fragments consisting of the VL, VH, CL and CH1 domains; a F (ab') 2 fragment which is a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; an Fd fragment consisting of the VH and CH1 domains; (iii) an Fv fragment consisting of VL and VH domains, a single chain Fv fragment (scFv) consisting of VL and VH domains connected by a linker sequence; and single domain antibody (dAb) fragments consisting of either a VH domain or a VL domain (Ward et al, 1989Nature [ Nature ]341: 544-546).

The term "single chain antibody", "single chain Fv" or "scFv" means a molecule comprising an antibody heavy chain variable domain (or region; VH) and an antibody light chain variable domain (or region; VL) joined by a linker. Such scFv molecules can have the general structure: NH 2-VL-linker-VH-COOH or NH 2-VH-linker-VL-COOH. Any suitable linker may be used. A non-limiting set of linkers that can be used in such single chain antibodies is described in the following documents: holliger et al (1993), Proc. Natl. Acad. Sci. USA [ Proc. Natl. Acad. Sci. ]90: 6444-; the contents of each are hereby incorporated by reference. The term "antigen-binding fragment" of an antibody is also intended to encompass such single chain antibodies. These antibody fragments are obtained using techniques known to those skilled in the art and are screened for efficacy in the same manner as intact antibodies. Without limitation, antigen-binding fragments may be produced by any suitable method known in the art. For example, the various antigen-binding fragments described herein can be generated by enzymatic or chemical modification of whole antibodies, synthesized de novo using recombinant DNA methods (e.g., single chain Fv), or identified using phage display libraries (see, e.g., Pini and Bracci, Curr Protein Pept Sci [ Current Protein and peptide sciences ] 2000; 1(2):155-69, the contents of which are hereby incorporated by reference). Antigen binding fragments are screened for utility (e.g., binding affinity, activity) in the same manner as intact antibodies.

Antigen-binding fragments may also be incorporated into single domain antibodies, large antibodies (maxibodies), minibodies (minibodies), intrabodies, diabodies, triabodies, tetrabodies, v-NARs, and bis-scFvs (see, e.g., Hollinger and Hudson,2005, Nature Biotechnology [ Nature Biotechnology ],23,9,1126-1136, the contents of which are hereby incorporated by reference). The antigen-binding portion of the antibody can be grafted into a scaffold based on a polypeptide such as fibronectin type III (Fn3) (see, e.g., U.S. patent No. 6,703,199, which describes fibronectin polypeptide monomers, the contents of which are hereby incorporated by reference).

The antigen-binding fragment may be incorporated into a single chain molecule comprising a pair of tandem Fv segments (VH-CH1-VH-CH1) that together with a complementary light chain polypeptide form a pair of antigen-binding regions (see Zapata et al, 1995Protein Eng. [ Protein engineering ]8(10): 1057-1062; and U.S. Pat. No. 5,641,870; the contents of each of which are hereby incorporated by reference).

Throughout the specification, the term "isolated" means that an immunoglobulin, antibody or polynucleotide (as the case may be) is present in a physical environment different from that in the natural environment. For example, a naturally occurring polynucleotide or polypeptide present in a living organism is not isolated, but the same polynucleotide or polypeptide that is separated from some or all of the coexisting materials in the living organism is isolated.

As used herein, the term "isolated antibody" refers to an antibody that has been identified and separated from one or more (e.g., a majority) of the components (by weight) in its environment of origin, e.g., from components of a hybridoma cell culture or a different cell culture for the production thereof (e.g., a producer cell, including but not limited to the exemplary host cell described herein that recombinantly expresses the antibody). The separation is performed such that it substantially removes components that may otherwise interfere with the suitability of the antibody for the desired application (e.g., therapeutic use against an anti-NPR 1 antibody). Methods of preparing isolated antibodies are known in the art and include protein a chromatography, anion exchange chromatography, cation exchange chromatography, virus retention filtration, and ultrafiltration.

Throughout the specification, complementarity determining regions ("CDRs") are defined according to the kabat definition, unless it is stated that CDRs are defined according to another definition. The precise amino acid sequence boundaries of a given CDR can be determined using any of a number of well-known schemes, including those described below: kabat et al (1991), "Sequences of Proteins of Immunological Interest". 5 th edition Public Health Service,. American National Institutes of Health (National Institutes of Health), Bethesda, Maryland (MD) ("Kabat" numbering scheme); Al-Lazikani et Al, (1997) JMB 273,927-948 ("Georgia" numbering scheme) and ImmunoGenTics (IMGT) numbering (Lefranc, M. -P., The Immunogligst [ Immunologist ],7,132-136 (1999); Lefranc, M. -P. et Al, Dev. Comp. Immunol. [ developmental and comparative immunology ],27,55-77(2003) ("IMGT" numbering scheme); The respective contents are hereby incorporated by reference for The classical forms, for example, CDR amino acid residues in The heavy chain variable domain (VH) are numbered 31-35(HCDR1), 50-65(HCDR2) and 95-102(HCDR3) according to Carbart, CDR amino acid residues in The light chain variable domain (VL) are numbered 24-34(LCDR1), 50-56(LCDR2) and 56(HCDR 4682) are numbered according to Calgarland 25-5926, 52-56(HCDR2) and 95-102(HCDR 3); and amino acid residues in VL are numbered 26-32(LCDR1), 50-52(LCDR2) and 91-96(LCDR 3). By combining the CDR definitions of both kabat and georgia, the CDRs consist of amino acid residues 26-35(HCDR1), 50-65(HCDR2) and 95-102(HCDR3) in the human VH and amino acid residues 24-34(LCDR1), 50-56(LCDR2) and 89-97(LCDR3) in the human VL. According to IMGT, the CDR amino acid residues in the VH are numbered approximately 26-35(CDR1), 51-57(CDR2) and 93-102(CDR3), and the CDR amino acid residues in the VL are numbered approximately 27-32(CDR1), 50-52(CDR2) and 89-97(CDR3) (numbered according to "kabat"). Under IMGT, the program IMGT/DomainGap Align can be used to determine the CDR regions of antibodies.

By convention, the CDR regions in the heavy chain are commonly referred to as HCDR1, HCDR2 and HCDR3, and the CDR regions in the light chain are commonly referred to as LCDR1, LCDR2 and LCDR 3. They are numbered sequentially in the direction from the amino terminus to the carboxy terminus.

As used herein, the term "antibody framework" refers to a portion of a variable domain, i.e., VL or VH, that serves as a scaffold for the antigen binding loops (CDRs) of the variable domain. Essentially, it is a variable domain without CDRs.

The term "constant region" or "constant domain" refers to the carboxy-terminal portion of the light and heavy chains that are not directly involved in binding of the antibody to the antigen, but exhibit multiple effector functions, such as interaction with an Fc receptor. The term refers to a portion of an immunoglobulin molecule that has a more conserved amino acid sequence relative to another portion of an immunoglobulin, i.e., a variable domain, and comprises an antigen binding site. The constant domains comprise the CH1, CH2, and CH3 domains of the heavy chain, and the CHL domain of the light chain.

The term "epitope" or "antigenic determinant" refers to a site on an antigen to which an immunoglobulin or antibody specifically binds (e.g., a specific site on a target molecule). Epitopes typically comprise at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acids, conserved or not, in a unique spatial conformation. See, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, volume 66, edited by g.e. morris (1996), the contents of which are hereby incorporated by reference. In addition, as used herein, an epitope may comprise one or more monosaccharide units of the polysaccharide to which the antibody specifically binds. In particular aspects, the epitope can be a conformational epitope. See, e.g., Thompson et al, 2009, J.of biol.chem. [ J.Biol.Chem. ]51:35621-35631, the contents of which are hereby incorporated by reference.

As used herein, the term "monoclonal antibody" or "monoclonal antibody composition" refers to an antibody obtained from a substantially homogeneous population of antibodies produced by a particular cell or cell line, wherein the individual antibodies comprising the population are substantially identical in sequence, except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibody preparations exhibit a single binding specificity and affinity for a particular epitope. In contrast, conventional (polyclonal) antibody preparations typically include multiple antibodies directed against or specific for different epitopes. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. Monoclonal antibodies (mAbs) can be produced by a variety of techniques including conventional methods such as standard somatic hybridization techniques of Kohler and Milstein (Nature [ Nature ] 1975; 256(5517):495-7, the contents of which are hereby incorporated by reference). Monoclonal antibodies can also be obtained from other suitable methods, including phage display techniques, such as those described in the following references: clackson et al (Nature [ Nature ] 1991; 352(6336):624-8) or Marks et al (J Mol Biol [ journal of molecular biology ] 1991; 222(3):581-97), the contents of each of which are hereby incorporated by reference. The term "monoclonal antibody" is also not limited to antibody sequences from a particular source species or from a single source species. Thus, the term "monoclonal antibody" is meant to encompass chimeric monoclonal antibodies, such as humanized monoclonal antibodies.

As used herein, the term "chimeric antibody" refers to an antibody in which (a) the constant regions are altered, replaced, or exchanged such that the antigen binding site (variable region) is linked to a constant region that is different or altered in class, effector function, and/or species; or (b) altering, replacing or exchanging the variable region or portion thereof with a variable region or portion thereof having a different or altered antigenic specificity. To generate chimeric antibodies, variable region sequences from a non-human donor antibody (e.g., a mouse, rabbit, or rat donor antibody) can be linked to human constant regions using methods known in the art (see, e.g., U.S. Pat. No. 4,816,567(Cabilly et al), the contents of which are hereby incorporated by reference). For example, a mouse anti-NPR 1 antibody can be modified by replacing its constant region with a constant region from a human immunoglobulin. Due to the replacement by human constant regions, the chimeric antibody can retain its specificity of recognizing human NPR1 while having reduced immunogenicity in humans compared to the original mouse antibody.

The term "humanized antibody" as used herein refers to a form of an antibody that comprises at least some human sequences and at least some non-human sequences. Typically, antibodies comprise a human sequence and a small portion of a non-human sequence that confers binding specificity to a target antigen. Such antibodies are chimeric antibodies that contain minimal sequences derived from non-human immunoglobulins and retain the reactivity of the non-human antibodies while being less immunogenic in humans. Typically, humanized antibodies are generated by replacing the hypervariable region sequences of a human recipient antibody with a non-human donor antibody (e.g., a mouse, rabbit or rat donor antibody) that binds to the antigen of interest (e.g., NPR 1). In some cases, the framework region sequences of the acceptor antibody may also be replaced with the corresponding sequences of the donor antibody (e.g., via affinity maturation). In addition to sequences derived from donor and acceptor antibodies, humanized antibodies may be modified by residue substitutions in the framework regions and/or substituted non-human residues to improve and optimize antibody specificity, affinity, and/or activity as described herein. Methods of producing humanized antibodies are known in the art. See, e.g., Riechmann et al (Nature [ Nature ] 1988; 332(6162): 323-7); jones et al (Nature [ Nature ] 1986; 321(6069): 522-5); U.S. Pat. No. 5,225,539 (Winter); and U.S. Pat. Nos. 5,530,101, 5,585,089, 5,693,762, and 6,180,370(Queen et al), the contents of each of which are hereby incorporated by reference.

As used herein, the term "human antibody" is intended to include antibodies having variable regions in which both the framework and CDR regions are derived from human-derived sequences. Furthermore, if the antibody contains constant regions, the constant regions are also derived from such human sequences, e.g., human germline sequences, or mutated versions of human germline sequences or antibodies containing consensus framework sequences derived from analysis of human framework sequences, e.g., as described by Knappik et al, (2000), J Mol Biol [ journal of molecular biology ]; 296:57-86, the contents of which are hereby incorporated by reference. Human antibodies can include amino acid residues that are not encoded by human sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo).

The antibodies or antigen-binding fragments of the disclosure may include amino acid residues that are not encoded by human sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species (e.g., a mouse) have been grafted into human framework sequences.

The terms "peptide," "polypeptide," and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms encompass amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of the corresponding naturally occurring amino acid, as well as apply to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. Unless otherwise indicated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.

The term "conservatively modified variants" applies to both amino acid and nucleic acid sequences. With respect to nucleic acid sequences, conservatively modified variants refers to those nucleic acids which encode identical or essentially identical amino acid sequences, or where the nucleic acid does not encode an amino acid sequence, to essentially identical sequences. Due to the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For example, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at each position where an alanine is specified by a codon, the codon can be changed to any of the corresponding codons described without changing the encoded polypeptide. Such nucleic acid variations are "silent variations," which are one of the conservatively modified variations. Every nucleic acid sequence herein that encodes a polypeptide also describes every possible silent variation of the nucleic acid. The skilled artisan will recognize that each codon in a nucleic acid (except AUG, which is typically the only codon for methionine, and TGG, which is typically the only codon for tryptophan) can be modified to produce a functionally identical molecule. Thus, each silent variation of a nucleic acid encoding a polypeptide is implicit in each such sequence. With respect to polypeptide sequences, "conservatively modified variants" includes single substitutions, deletions or additions to a polypeptide sequence such that an amino acid is substituted with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. The following group 8 contains amino acids that are conservative substitutions for each other:

1) Alanine (a), glycine (G);

2) aspartic acid (D), glutamic acid (E);

3) asparagine (N), glutamine (Q);

4) arginine (R), lysine (K);

5) isoleucine (I), leucine (L), methionine (M), valine (V);

6) phenylalanine (F), tyrosine (Y), tryptophan (W);

7) serine (S), threonine (T); and

8) cysteine (C), methionine (M).

The term "identity" or "homology" refers to the relationship between the sequences of two or more polypeptides as determined by comparing the sequences. "identity" also refers to the degree of sequence relatedness between polypeptides, as determined by the number of matches between strings of two or more amino acid residues. The percent identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., percent identity equals the number of identical positions/total number of positions x 100), which, taking into account the number of gaps and the length of each gap, need to be introduced in order to optimally align the two sequences. Sequence comparison and percent identity determination between two sequences can be accomplished using a mathematical algorithm. For sequence comparison, typically one sequence acts as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, the test sequence and the reference sequence are input into a computer, subsequence coordinates are designated as necessary, and sequence algorithm program parameters are designated. Default program parameters may be used, or alternative parameters may be specified. The sequence comparison algorithm will then calculate the percent sequence identity of the test sequence relative to the reference sequence based on the program parameters. Additionally or alternatively, protein sequences of the disclosure can be further used as "query sequences" to search public databases, for example, to identify related sequences. Such searches may be performed, for example, using the BLAST program of Altschul et al (J Mol Biol 1990; 215(3):403-10, the contents of which are hereby incorporated by reference).

Two sequences are "substantially identical" if they have a specified percentage of amino acid residues or nucleotides that are identical (e.g., 60% identity, 65% identity, 70% identity, 75% identity, 80% identity, 85% identity, 90% identity, 95% identity, or 99% identity over a specified region or over the entire sequence when not specified) when compared and aligned over a comparison window or designated region to obtain the maximum correspondence when measured using one of the following sequence comparison algorithms or by manual alignment and visual inspection. Optionally, the identity is present over a region of at least about 50 nucleotides (or 10 amino acids) in length, or over a region of 100 to 500 or 1000 or more nucleotides (or 20, 50, 200 or more amino acids) in length.

Binding "affinity" refers to the strength of interaction between an antibody and an antigen at a single point of antigen localization. Within each antigenic site, the variable region of the antibody "arm" interacts with the antigen at many sites through weak non-covalent forces. In general, the more interactions, the stronger the affinity. Typically, such determinations can be made using cell-based assays.

As used herein, the term "Kassoc" or "Ka" is intended to refer to the association rate of a particular binding molecule-antigen interaction, while the term "Kdis" or "Kd" as used herein is intended to refer to the dissociation rate of a particular binding molecule-antigen interaction. As used herein, the term "K_D"is intended to mean the equilibrium dissociation constant, which is obtained from the ratio of Kd to Ka (i.e. Kd/Ka) and expressed as molar concentration (M). The K of an antibody can be determined using well established methods in the art_DThe value is obtained. Determination of K of antibodies_DBy using surface plasmon resonance (e.g. using

Systematic or Solution Equilibrium Titration (SET)) was performed (see Friguet et al (1985) j. immunol. methods [ journal of immunological methods ]](ii) a 77(2), 305-319; and Hanel et al (2005) anal. biochem. [ analytical biochemistry ]](ii) a 339(1) 182, 184, the respective contents of which are hereby incorporated by reference.

As used herein, the terms "specific", "specific binding" and "specific binding" refer to a binding reaction between an antibody or antigen-binding fragment (e.g., an anti-NPR 1 antibody) and a target antigen (e.g., NPR1) in a homogeneous population of proteins and other biologicals. The binding specificity of an antibody can be tested by comparing binding to the appropriate antigen and binding to an unrelated antigen or mixture of antigens under a given set of conditions. An antibody is considered specific if it binds to the appropriate antigen with at least 2, 5, 7, preferably 10 or more times higher affinity than the irrelevant antigen or antigen mixture. A "specific antibody" or "target-specific antibody" is an antibody that binds only to (or exhibits minimal binding to) a target antigen (e.g., NPR1) but not to other antigens Combined). In certain embodiments, the K of an antibody or antigen-binding fragment that specifically binds a target antigen (e.g., NPR1)_DLess than 1x10^-6M, less than 1x10^-7M, less than 1x10^-8M, less than 1x10^-9M, less than 1x10^-10M, less than 1x10^-11M, less than 1x10^-12M or less than 1x10^-13And M. In certain embodiments, K_DFrom about 1pM to about 600 pM. In certain embodiments, K_DBetween 600pM to 1. mu.M, 1. mu.M to 100nM or 100mM to 10nM, inclusive.

In some embodiments, the antibody or antigen-binding fragment thereof acts as a non-competitive agonist. "non-competitive agonist" refers to a molecule that binds to an enzyme or receptor at a site remote from the binding site of its natural ligand. Noncompetitive or allosteric agonists are generally independent of the association or concentration of the enzyme or the natural ligand of the receptor. Such non-competitive agonists may, for example, provide a level of activation that may be substantially independent of the native ligand. In a specific embodiment, the anti-NPR 1 antibody or antigen-binding fragment described herein is ANP non-competitive, meaning that the antibody or antigen-binding fragment acts as an agonist, binding at a site distant from the ANP binding site of NPR1 and affecting agonist activity, regardless of whether NPR1 binds to ANP.

In some embodiments, the antibody or antigen-binding fragment thereof acts as a competitive agonist. "competitive agonist" refers to an agonist that interferes with or competes with a natural ligand for its binding site on an enzyme or receptor. In a specific embodiment, the anti-NPR 1 antibody or antigen-binding fragment described herein is ANP-competitive, meaning that the antibody or antigen-binding fragment acts as an agonist that competes with ANP at the ANP-binding site of NPR 1.

In some embodiments, activation of NPR1 by the antibody or antigen-binding fragment can be determined by any suitable assay. An exemplary assay for determining NPR1 activation is cGMP production from a mammalian cell (e.g., CHO cell or human cell line) expressing hNPR 1.

In some embodiments, stabilization of the ANP-NPR1 complex may be determined by any suitable assay. An exemplary assay for determining the stability of the ANP-NPR1 complex is the FRET assay described herein (see, e.g., fig. 3).

The term "about" with respect to the numerical value x means, for example, x ± 10%.

Antibodies of the disclosure

Certain specific anti-NPR 1 antibody sequences of the present disclosure are disclosed below. As used herein, the term "anti-NPR 1 antibody" or "antibody that binds NPR 1" refers to any form of antibody or antigen-binding fragment that specifically binds NPR1, e.g., at less than 1x10 ^-8K of M_DThose that bind, as determined by, for example, Surface Plasmon Resonance (SPR) spectroscopy (using Biacore)^TM) Or Solution Equilibrium Titration (SET). The term includes monoclonal antibodies (including intact monoclonal antibodies), polyclonal antibodies, and biologically functional antigen binding fragments, so long as they specifically bind NPR 1.

Table 2 lists the amino acid and nucleic acid sequences of exemplary anti-NPR 1 antibodies of the disclosure. In some embodiments, the antibody has the heavy and light chain CDR, VH and VL sequences and/or the heavy and light chain sequences of any of the antibodies described in table 2. In some embodiments, the anti-NPR 1 antibody is a four chain antibody (also referred to as a whole antibody) comprising two heavy chains and two light chains. In some embodiments, the anti-NPR 1 antibody is an antigen-binding fragment of an intact antibody, e.g., a functional fragment of an intact antibody selected from those listed in table 2, which retains the ability to bind NPR1 and/or provide intact antibody function (e.g., activate NPR1 in the absence of ANP). In some embodiments, the anti-NPR 1 antibody is an antibody having the CDRs of any of the heavy chain variable region and light chain variable region pairs shown in table 2. In some embodiments, the anti-NPR 1 antibody is an antibody having the CDRs of any of the heavy and light chain pairs shown in table 2.

TABLE 2 exemplary anti-NPR 1 antibody sequences

For example, WW01_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) SEQ ID NO 4(HCDR1), SEQ ID NO 5(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 5(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 9(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 11(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3). In some embodiments, WW01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 4(HCDR1), SEQ ID NO 5(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, WW01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 7(HCDR1), SEQ ID NO 5(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, WW01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 8(HCDR1), SEQ ID NO 9(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3). In some embodiments, WW01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 10(HCDR1), SEQ ID NO 11(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, WW01_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 13 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 24. In some embodiments, WW01_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 15 and a light chain comprising or having the amino acid sequence of SEQ ID No. 26.

For example, WW03_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 29(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 43(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:43(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:46(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 43(LCDR 3). In some embodiments, WW03_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 43(LCDR 3). In some embodiments, WW03_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:43(LCDR 3). In some embodiments, WW03_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:46(LCDR 3). In some embodiments, WW03_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:35(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:43(LCDR 3). In some embodiments, WW03_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 48. In some embodiments, WW03_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 39 and a light chain comprising or having the amino acid sequence of SEQ ID No. 50.

For example, WW05_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 52(HCDR1), 53(HCDR2), 54(HCDR3), 65(LCDR1), 66(LCDR2) and 67(LCDR3) SEQ ID NO; (II) SEQ ID NO:55(HCDR1), SEQ ID NO:53(HCDR2), SEQ ID NO:54(HCDR3), SEQ ID NO:65(LCDR1), SEQ ID NO:66(LCDR2) and SEQ ID NO:67(LCDR 3); (III) SEQ ID NO:56(HCDR1), SEQ ID NO:57(HCDR2), SEQ ID NO:54(HCDR3), SEQ ID NO:68(LCDR1), SEQ ID NO:69(LCDR2), and SEQ ID NO:70(LCDR 3); or (IV) SEQ ID NO 58(HCDR1), SEQ ID NO 59(HCDR2), SEQ ID NO 60(HCDR3), SEQ ID NO 71(LCDR1), SEQ ID NO 69(LCDR2) and SEQ ID NO 67(LCDR 3). In some embodiments, WW05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:52(HCDR1), SEQ ID NO:53(HCDR2), SEQ ID NO:54(HCDR3), SEQ ID NO:65(LCDR1), SEQ ID NO:66(LCDR2), and SEQ ID NO:67(LCDR 3). In some embodiments, WW05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:55(HCDR1), SEQ ID NO:53(HCDR2), SEQ ID NO:54(HCDR3), SEQ ID NO:65(LCDR1), SEQ ID NO:66(LCDR2), and SEQ ID NO:67(LCDR 3). In some embodiments, WW05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:56(HCDR1), SEQ ID NO:57(HCDR2), SEQ ID NO:54(HCDR3), SEQ ID NO:68(LCDR1), SEQ ID NO:69(LCDR2), and SEQ ID NO:70(LCDR 3). In some embodiments, WW05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:58(HCDR1), SEQ ID NO:59(HCDR2), SEQ ID NO:60(HCDR3), SEQ ID NO:71(LCDR1), SEQ ID NO:69(LCDR2), and SEQ ID NO:67(LCDR 3). In some embodiments, WW05_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 61 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 72. In some embodiments, WW05_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 63 and a light chain comprising or having the amino acid sequence of SEQ ID No. 74.

For example, WW06_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 76(HCDR1) SEQ ID NO 77(HCDR2), 78(HCDR3), 89(LCDR1), 90(LCDR2) and 91(LCDR3) SEQ ID NO; (II) SEQ ID NO:79(HCDR1), SEQ ID NO:77(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:89(LCDR1), SEQ ID NO:90(LCDR2) and SEQ ID NO:91(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:81(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:92(LCDR1), SEQ ID NO:93(LCDR2), and SEQ ID NO:94(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 83(HCDR2), SEQ ID NO 84(HCDR3), SEQ ID NO 95(LCDR1), SEQ ID NO 93(LCDR2) and SEQ ID NO 91(LCDR 3). In some embodiments, WW06_ LALA may be defined as comprising or having the amino acid sequence: 76(HCDR1) SEQ ID NO 77(HCDR2) SEQ ID NO 78(HCDR3), 89(LCDR1) SEQ ID NO 90(LCDR2) and 91(LCDR 3). In some embodiments, WW06_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:79(HCDR1), SEQ ID NO:77(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:89(LCDR1), SEQ ID NO:90(LCDR2), and SEQ ID NO:91(LCDR 3). In some embodiments, WW06_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:80(HCDR1), SEQ ID NO:81(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:92(LCDR1), SEQ ID NO:93(LCDR2), and SEQ ID NO:94(LCDR 3). In some embodiments, WW06_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:82(HCDR1), SEQ ID NO:83(HCDR2), SEQ ID NO:84(HCDR3), SEQ ID NO:95(LCDR1), SEQ ID NO:93(LCDR2), and SEQ ID NO:91(LCDR 3). In some embodiments, WW06_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:85 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 96. In some embodiments, WW06_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 87 and a light chain comprising or having the amino acid sequence of SEQ ID No. 98.

For example, XX01_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) SEQ ID NO 4(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 101(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 4(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 8(HCDR1), SEQ ID NO 101(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3). In some embodiments, XX01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 10(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 103 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 24. In some embodiments, XX01_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 105 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 26.

For example, XX01_ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) SEQ ID NO 4(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 101(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 4(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 8(HCDR1), SEQ ID NO 101(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3). In some embodiments, XX01_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 10(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO. 103 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO. 24. In some embodiments, XX01_ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO 108 and a light chain comprising or having the amino acid sequence of SEQ ID NO 26.

For example, XX01_ N30S _ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 112(HCDR1), 100(HCDR2), 6(HCDR3), 17(LCDR1), 18(LCDR2) and 19(LCDR3) SEQ ID NO; (II) SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 113(HCDR1), SEQ ID NO 101(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 114(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ N30S _ DAPA may be defined as comprising or having the amino acid sequence: 112(HCDR1), 100(HCDR2), 6(HCDR3), 17(LCDR1), 18(LCDR2), and 19(LCDR 3). In some embodiments, XX01_ N30S _ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ N30S _ DAPA may be defined as comprising or having the amino acid sequence: 113(HCDR1), 101(HCDR2), 6(HCDR3), 20(LCDR1), 21(LCDR2), and 22(LCDR 3). In some embodiments, XX01_ N30S _ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 114(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX01_ N30S _ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO 115 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO 24. In some embodiments, XX01_ N30S _ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO 117 and a light chain comprising or having the amino acid sequence of SEQ ID NO 26.

For example, XX03_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 43(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:43(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:46(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 43(LCDR 3). In some embodiments, XX03_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 43(LCDR 3). In some embodiments, XX03_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:43(LCDR 3). In some embodiments, XX03_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:46(LCDR 3). In some embodiments, XX03_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:43(LCDR 3). In some embodiments, XX03_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 48. In some embodiments, XX03_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 124 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 50.

For example, XX04_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 29(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 126(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:126(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 126(LCDR 3). In some embodiments, XX04_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 126(LCDR 3). In some embodiments, XX04_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:126(LCDR 3). In some embodiments, XX04_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3). In some embodiments, XX04_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:35(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:126(LCDR 3). In some embodiments, XX04_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 128. In some embodiments, XX04_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 39 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 130.

For example, XX06_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 29(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 134(LCDR 3). In some embodiments, XX06_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 134(LCDR 3). In some embodiments, XX06_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX06_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:135(LCDR 3). In some embodiments, XX06_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:35(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX06_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 136. In some embodiments, XX06_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 39 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 138.

For example, XX06_ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 29(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 134(LCDR 3). In some embodiments, XX06_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 134(LCDR 3). In some embodiments, XX06_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX06_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:135(LCDR 3). In some embodiments, XX06_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:35(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX06_ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 136. In some embodiments, XX06_ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 141 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 138.

For example, XX07_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 29(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 145(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:145(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:146(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 145(LCDR 3). In some embodiments, XX07_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 145(LCDR 3). In some embodiments, XX07_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:145(LCDR 3). In some embodiments, XX07_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:146(LCDR 3). In some embodiments, XX07_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:35(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:145(LCDR 3). In some embodiments, XX07_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 147. In some embodiments, XX07_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 39 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 149.

For example, XX08_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) SEQ ID NO 4(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 4(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 8(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3). In some embodiments, XX08_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 10(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:154 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 24. In some embodiments, XX08_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 156 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 26.

For example, XX08_ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) SEQ ID NO 4(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 4(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 8(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 22(LCDR 3). In some embodiments, XX08_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 10(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:154 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 24. In some embodiments, XX08_ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 159 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 26.

For example, XX08_ N30S _ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 112(HCDR1) SEQ ID NO 151(HCDR2), 6(HCDR3), 17(LCDR1), 18(LCDR2) and 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 113(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 114(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ N30S _ DAPA may be defined as comprising or having the amino acid sequence: 112(HCDR1) SEQ ID NO 151(HCDR2), 6(HCDR3), 17(LCDR1), 18(LCDR2) and 19(LCDR 3). In some embodiments, XX08_ N30S _ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ N30S _ DAPA may be defined as comprising or having the amino acid sequence: 113(HCDR1), 152(HCDR2), 6(HCDR3), 20(LCDR1), 21(LCDR2), and 22(LCDR 3). In some embodiments, XX08_ N30S _ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 114(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ N30S _ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO. 161 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO. 24. In some embodiments, XX08_ N30S _ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO 163 and a light chain comprising or having the amino acid sequence of SEQ ID NO 26.

For example, XX08_ N30Q _ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 165(HCDR1) SEQ ID NO, 151(HCDR2) SEQ ID NO 6(HCDR3), 17(LCDR1) SEQ ID NO 18(LCDR2) and 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 166(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO:167(HCDR1), SEQ ID NO:153(HCDR2), SEQ ID NO:12(HCDR3), SEQ ID NO:23(LCDR1), SEQ ID NO:21(LCDR2) and SEQ ID NO:19(LCDR 3). In some embodiments, XX08_ N30Q _ DAPA may be defined as comprising or having the amino acid sequence: 165(HCDR1) SEQ ID NO, 151(HCDR2) SEQ ID NO 6(HCDR3), 17(LCDR1) SEQ ID NO 18(LCDR2) and 19(LCDR 3). In some embodiments, XX08_ N30Q _ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2), and SEQ ID NO 19(LCDR 3). In some embodiments, XX08_ N30Q _ DAPA may be defined as comprising or having the amino acid sequence: 166(HCDR1) SEQ ID NO 152(HCDR2), 6(HCDR3), 20(LCDR1), 21(LCDR2), and 22(LCDR 3). In some embodiments, XX08_ N30Q _ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:167(HCDR1), SEQ ID NO:153(HCDR2), SEQ ID NO:12(HCDR3), SEQ ID NO:23(LCDR1), SEQ ID NO:21(LCDR2), and SEQ ID NO:19(LCDR 3). In some embodiments, XX08_ N30Q _ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:168 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 24. In some embodiments, XX08_ N30Q _ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO:170 and a light chain comprising or having the amino acid sequence of SEQ ID NO: 26.

For example, XX09_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 29(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 172(LCDR3) SEQ ID NO; (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 172(LCDR 3). In some embodiments, XX09_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 172(LCDR 3). In some embodiments, XX09_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:172(LCDR 3). In some embodiments, XX09_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:173(LCDR 3). In some embodiments, XX09_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:35(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:172(LCDR 3). In some embodiments, XX09_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 174. In some embodiments, XX09_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 39 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 176.

For example, XX11_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 29(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 178(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:178(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 178(LCDR 3). In some embodiments, XX11_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 178(LCDR 3). In some embodiments, XX11_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:178(LCDR 3). In some embodiments, XX11_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3). In some embodiments, XX11_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:35(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:178(LCDR 3). In some embodiments, XX11_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 180. In some embodiments, XX11_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 39 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 182.

For example, XX12_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 29(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 184(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:184(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:185(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 184(LCDR 3). In some embodiments, XX12_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 184(LCDR 3). In some embodiments, XX12_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:184(LCDR 3). In some embodiments, XX12_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:185(LCDR 3). In some embodiments, XX12_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:35(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:184(LCDR 3). In some embodiments, XX12_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 186. In some embodiments, XX12_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 39 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 188.

For example, XX13_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1), 190(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:190(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:191(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 192(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 134(LCDR 3). In some embodiments, XX13_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 190(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 134(LCDR 3). In some embodiments, XX13_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:190(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX13_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:191(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:135(LCDR 3). In some embodiments, XX13_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:192(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX13_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 193 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 136. In some embodiments, XX13_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 195 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 138.

For example, XX14_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 190(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 172(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:190(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:191(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 192(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 172(LCDR 3). In some embodiments, XX14_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 190(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 172(LCDR 3). In some embodiments, XX14_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:190(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:172(LCDR 3). In some embodiments, XX14_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:191(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:173(LCDR 3). In some embodiments, XX14_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:192(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:172(LCDR 3). In some embodiments, XX14_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 193 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 174. In some embodiments, XX14_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 195 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 176.

For example, XX15_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 126(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:126(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 126(LCDR 3). In some embodiments, XX15_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 126(LCDR 3). In some embodiments, XX15_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:126(LCDR 3). In some embodiments, XX15_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3). In some embodiments, XX15_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:126(LCDR 3). In some embodiments, XX15_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 128. In some embodiments, XX15_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 203 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 130.

For example, XX15_ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 126(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:126(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 126(LCDR 3). In some embodiments, XX15_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 126(LCDR 3). In some embodiments, XX15_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:126(LCDR 3). In some embodiments, XX15_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3). In some embodiments, XX15_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:126(LCDR 3). In some embodiments, XX15_ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO. 128. In some embodiments, XX15_ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO 208 and a light chain comprising or having the amino acid sequence of SEQ ID NO 130.

For example, XX16_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 134(LCDR 3). In some embodiments, XX16_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 134(LCDR 3). In some embodiments, XX16_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX16_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:135(LCDR 3). In some embodiments, XX16_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX16_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 136. In some embodiments, XX16_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 203 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 138.

For example, XX16_ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 134(LCDR 3). In some embodiments, XX16_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 134(LCDR 3). In some embodiments, XX16_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX16_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:135(LCDR 3). In some embodiments, XX16_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:134(LCDR 3). In some embodiments, XX16_ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO. 136. In some embodiments, XX16_ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO 208 and a light chain comprising or having the amino acid sequence of SEQ ID NO 138.

For example, XX17_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 145(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:145(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:146(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 145(LCDR 3). In some embodiments, XX17_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 145(LCDR 3). In some embodiments, XX17_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:145(LCDR 3). In some embodiments, XX17_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:146(LCDR 3). In some embodiments, XX17_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:145(LCDR 3). In some embodiments, XX17_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 147. In some embodiments, XX17_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 203 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 149.

For example, XX17_ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 145(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:145(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:146(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 145(LCDR 3). In some embodiments, XX17_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 145(LCDR 3). In some embodiments, XX17_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:145(LCDR 3). In some embodiments, XX17_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:146(LCDR 3). In some embodiments, XX17_ DAPA may be defined as comprising or having the amino acid sequence: (IV) SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:145(LCDR 3). In some embodiments, XX17_ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO. 147. In some embodiments, XX17_ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO 208 and a light chain comprising or having the amino acid sequence of SEQ ID NO 149.

For example, XX18_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 172(LCDR3) SEQ ID NO; (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 172(LCDR 3). In some embodiments, XX18_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 172(LCDR 3). In some embodiments, XX18_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:172(LCDR 3). In some embodiments, XX18_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:173(LCDR 3). In some embodiments, XX18_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:172(LCDR 3). In some embodiments, XX18_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 174. In some embodiments, XX18_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 203 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 176.

For example, XX18_ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 172(LCDR3) SEQ ID NO; (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 172(LCDR 3). In some embodiments, XX18_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 172(LCDR 3). In some embodiments, XX18_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:172(LCDR 3). In some embodiments, XX18_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:173(LCDR 3). In some embodiments, XX18_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:172(LCDR 3). In some embodiments, XX18_ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO. 174. In some embodiments, XX18_ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO 208 and a light chain comprising or having the amino acid sequence of SEQ ID NO 176.

For example, XX19_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 178(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:178(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 178(LCDR 3). In some embodiments, XX19_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 178(LCDR 3). In some embodiments, XX19_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:178(LCDR 3). In some embodiments, XX19_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3). In some embodiments, XX19_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:178(LCDR 3). In some embodiments, XX19_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 180. In some embodiments, XX19_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 203 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 182.

For example, XX19_ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 178(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:178(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 178(LCDR 3). In some embodiments, XX19_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 178(LCDR 3). In some embodiments, XX19_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:178(LCDR 3). In some embodiments, XX19_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3). In some embodiments, XX19_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:178(LCDR 3). In some embodiments, XX19_ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO. 180. In some embodiments, XX19_ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO 208 and a light chain comprising or having the amino acid sequence of SEQ ID NO 182.

For example, XX20_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 184(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:184(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:185(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 184(LCDR 3). In some embodiments, XX20_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 184(LCDR 3). In some embodiments, XX20_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:184(LCDR 3). In some embodiments, XX20_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:185(LCDR 3). In some embodiments, XX20_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:184(LCDR 3). In some embodiments, XX20_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 186. In some embodiments, XX20_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO. 203 and a light chain comprising or having the amino acid sequence of SEQ ID NO. 188.

For example, XX20_ DAPA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 28(HCDR1) SEQ ID NO 119(HCDR2), 30(HCDR3), 41(LCDR1), 42(LCDR2) and 184(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:184(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:185(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 184(LCDR 3). In some embodiments, XX20_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2), and SEQ ID NO 184(LCDR 3). In some embodiments, XX20_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2), and SEQ ID NO:184(LCDR 3). In some embodiments, XX20_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:185(LCDR 3). In some embodiments, XX20_ DAPA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:121(HCDR2), SEQ ID NO:36(HCDR3), SEQ ID NO:47(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:184(LCDR 3). In some embodiments, XX20_ DAPA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO. 186. In some embodiments, XX20_ DAPA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID NO 208 and a light chain comprising or having the amino acid sequence of SEQ ID NO 188.

For example, YY01_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 226(HCDR1), 227(HCDR2), 228(HCDR3), 237(LCDR1), 238(LCDR2) and 239(LCDR3) to SEQ ID NO; (II) SEQ ID NO:229(HCDR1), SEQ ID NO:227(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2) and SEQ ID NO:239(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:230(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 231(HCDR2), SEQ ID NO 232(HCDR3), SEQ ID NO 243(LCDR1), SEQ ID NO 241(LCDR2) and SEQ ID NO 239(LCDR 3). In some embodiments, YY01_ LALA may be defined as comprising or having the amino acid sequence: 226(HCDR1), 227(HCDR2), 228(HCDR3), 237(LCDR1), 238(LCDR2), and 239(LCDR 3). In some embodiments, YY01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:229(HCDR1), SEQ ID NO:227(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3). In some embodiments, YY01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:230(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3). In some embodiments, YY01_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:34(HCDR1), SEQ ID NO:231(HCDR2), SEQ ID NO:232(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:239(LCDR 3). In some embodiments, YY01_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 233 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 244. In some embodiments, YY01_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 235 and a light chain comprising or having the amino acid sequence of SEQ ID No. 246.

For example, YY02_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) SEQ ID NO:248(HCDR1), SEQ ID NO:249(HCDR2), SEQ ID NO:250(HCDR3), SEQ ID NO:260(LCDR1), SEQ ID NO:261(LCDR2), and SEQ ID NO:262(LCDR 3); (II) SEQ ID NO:251(HCDR1), SEQ ID NO:249(HCDR2), SEQ ID NO:250(HCDR3), SEQ ID NO:260(LCDR1), SEQ ID NO:261(LCDR2), and SEQ ID NO:262(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:252(HCDR2), SEQ ID NO:250(HCDR3), SEQ ID NO:263(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:264(LCDR 3); or (IV) SEQ ID NO:253(HCDR1), SEQ ID NO:254(HCDR2), SEQ ID NO:255(HCDR3), SEQ ID NO:265(LCDR1), SEQ ID NO:241(LCDR2) and SEQ ID NO:262(LCDR 3). In some embodiments, YY02_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:248(HCDR1), SEQ ID NO:249(HCDR2), SEQ ID NO:250(HCDR3), SEQ ID NO:260(LCDR1), SEQ ID NO:261(LCDR2), and SEQ ID NO:262(LCDR 3). In some embodiments, YY02_ LALA may be defined as comprising or having the amino acid sequence: 251(HCDR1) SEQ ID NO:249(HCDR2), 250(HCDR3), 260(LCDR1), 261(LCDR2), and 262(LCDR 3). In some embodiments, YY02_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:252(HCDR2), SEQ ID NO:250(HCDR3), SEQ ID NO:263(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:264(LCDR 3). In some embodiments, YY02_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:253(HCDR1), SEQ ID NO:254(HCDR2), SEQ ID NO:255(HCDR3), SEQ ID NO:265(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:262(LCDR 3). In some embodiments, YY02_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:256 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 266. In some embodiments, YY02_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 258 and a light chain comprising or having the amino acid sequence of SEQ ID No. 268.

For example, YY03_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 270(HCDR1), 271(HCDR2), 272(HCDR3), 282(LCDR1), 261(LCDR2) and 283(LCDR 3); (II) SEQ ID NO:273(HCDR1), SEQ ID NO:271(HCDR2), SEQ ID NO:272(HCDR3), SEQ ID NO:282(LCDR1), SEQ ID NO:261(LCDR2), and SEQ ID NO:283(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:274(HCDR2), SEQ ID NO:272(HCDR3), SEQ ID NO:284(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:285(LCDR 3); or (IV) SEQ ID NO 275(HCDR1), SEQ ID NO 276(HCDR2), SEQ ID NO 277(HCDR3), SEQ ID NO 286(LCDR1), SEQ ID NO 241(LCDR2) and SEQ ID NO 283(LCDR 3). In some embodiments, YY03_ LALA may be defined as comprising or having the amino acid sequence: 270(HCDR1), 271(HCDR2), 272(HCDR3), 282(LCDR1), 261(LCDR2), and 283(LCDR 3). In some embodiments, YY03_ LALA may be defined as comprising or having the amino acid sequence: 273(HCDR1) SEQ ID NO:271(HCDR2) SEQ ID NO:272(HCDR3), 282(LCDR1) SEQ ID NO:261(LCDR2) and 283(LCDR 3). In some embodiments, YY03_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:274(HCDR2), SEQ ID NO:272(HCDR3), SEQ ID NO:284(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:285(LCDR 3). In some embodiments, YY03_ LALA may be defined as comprising or having the amino acid sequence: 275(HCDR1), 276(HCDR2), 277(HCDR3), 286(LCDR1), 241(LCDR2), and 283(LCDR 3). In some embodiments, YY03_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:278 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 287. In some embodiments, YY03_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 280 and a light chain comprising or having the amino acid sequence of SEQ ID No. 289.

For example, YY04_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 291(HCDR1) SEQ ID NO, 292(HCDR2) SEQ ID NO, 293(HCDR3) SEQ ID NO, 237(LCDR1) SEQ ID NO, 238(LCDR2) and 304(LCDR 3); (II) SEQ ID NO:294(HCDR1), SEQ ID NO:292(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2) and SEQ ID NO:304(LCDR 3); (III) SEQ ID NO:295(HCDR1), SEQ ID NO:296(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:305(LCDR 3); or (IV) SEQ ID NO:297(HCDR1), SEQ ID NO:298(HCDR2), SEQ ID NO:299(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:304(LCDR 3). In some embodiments, YY04_ LALA may be defined as comprising or having the amino acid sequence: 291(HCDR1) SEQ ID NO, 292(HCDR2) SEQ ID NO, 293(HCDR3) SEQ ID NO, 237(LCDR1) SEQ ID NO, 238(LCDR2) and 304(LCDR 3). In some embodiments, YY04_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:294(HCDR1), SEQ ID NO:292(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:304(LCDR 3). In some embodiments, YY04_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:295(HCDR1), SEQ ID NO:296(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:305(LCDR 3). In some embodiments, YY04_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:297(HCDR1), SEQ ID NO:298(HCDR2), SEQ ID NO:299(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:304(LCDR 3). In some embodiments, YY04_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:300 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 306. In some embodiments, YY04_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 302 and a light chain comprising or having the amino acid sequence of SEQ ID No. 308.

For example, YY05_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 310(HCDR1), 311(HCDR2), 312(HCDR3), 320(LCDR1), 321(LCDR2) and 322(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:321(LCDR2), and SEQ ID NO:322(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:325(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 314(HCDR2), SEQ ID NO 315(HCDR3), SEQ ID NO 326(LCDR1), SEQ ID NO 324(LCDR2) and SEQ ID NO 322(LCDR 3). In some embodiments, YY05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:310(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:321(LCDR2), and SEQ ID NO:322(LCDR 3). In some embodiments, YY05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:321(LCDR2), and SEQ ID NO:322(LCDR 3). In some embodiments, YY05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:325(LCDR 3). In some embodiments, YY05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:82(HCDR1), SEQ ID NO:314(HCDR2), SEQ ID NO:315(HCDR3), SEQ ID NO:326(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:322(LCDR 3). In some embodiments, YY05_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:316 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 327. In some embodiments, YY05_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 318 and a light chain comprising or having the amino acid sequence of SEQ ID No. 329.

For example, YY06_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 270(HCDR1), 271(HCDR2), 331(HCDR3), 337(LCDR1), 338(LCDR2) and 339(LCDR3) SEQ ID NO; (II) SEQ ID NO:273(HCDR1), SEQ ID NO:271(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:274(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 275(HCDR1), SEQ ID NO 276(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3). In some embodiments, YY06_ LALA may be defined as comprising or having the amino acid sequence: 270(HCDR1), 271(HCDR2), 331(HCDR3), 337(LCDR1), 338(LCDR2), and 339(LCDR3) SEQ ID NO. In some embodiments, YY06_ LALA may be defined as comprising or having the amino acid sequence: 273(HCDR1) SEQ ID NO 271(HCDR2) SEQ ID NO 331(HCDR3), 337(LCDR1) SEQ ID NO 338(LCDR2) and 339(LCDR3) SEQ ID NO. In some embodiments, YY06_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:274(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3). In some embodiments, YY06_ LALA may be defined as comprising or having the amino acid sequence: 275(HCDR1), 276(HCDR2), 332(HCDR3), 343(LCDR1), 341(LCDR2), and 339(LCDR3) SEQ ID NO. In some embodiments, YY06_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 333 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 344. In some embodiments, YY06_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 335 and a light chain comprising or having the amino acid sequence of SEQ ID No. 346.

For example, YY07_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 310(HCDR1), 311(HCDR2), 348(HCDR3), 320(LCDR1), 354(LCDR2) and 355(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:355(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2) and SEQ ID NO:356(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 314(HCDR2), SEQ ID NO 349(HCDR3), SEQ ID NO 326(LCDR1), SEQ ID NO 324(LCDR2) and SEQ ID NO 355(LCDR 3). In some embodiments, YY07_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:310(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:355(LCDR 3). In some embodiments, YY07_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:355(LCDR 3). In some embodiments, YY07_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:356(LCDR 3). In some embodiments, YY07_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:82(HCDR1), SEQ ID NO:314(HCDR2), SEQ ID NO:349(HCDR3), SEQ ID NO:326(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:355(LCDR 3). In some embodiments, YY07_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:350 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 357. In some embodiments, YY07_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 352 and a light chain comprising or having the amino acid sequence of SEQ ID No. 359.

For example, ZZ05_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 310(HCDR1), 311(HCDR2), 348(HCDR3), 320(LCDR1), 354(LCDR2) and 361(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:361(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:362(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 314(HCDR2), SEQ ID NO 349(HCDR3), SEQ ID NO 326(LCDR1), SEQ ID NO 324(LCDR2) and SEQ ID NO 361(LCDR 3). In some embodiments, ZZ05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:310(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:361(LCDR 3). In some embodiments, ZZ05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:361(LCDR 3). In some embodiments, ZZ05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:362(LCDR 3). In some embodiments, ZZ05_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:82(HCDR1), SEQ ID NO:314(HCDR2), SEQ ID NO:349(HCDR3), SEQ ID NO:326(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:361(LCDR 3). In some embodiments, ZZ05_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:350 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 363. In some embodiments, ZZ05_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 352 and a light chain comprising or having the amino acid sequence of SEQ ID No. 365.

For example, ZZ12_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) SEQ ID NO:367(HCDR1), SEQ ID NO:368(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3); (II) SEQ ID NO 369(HCDR1), SEQ ID NO 368(HCDR2), SEQ ID NO 228(HCDR3), SEQ ID NO 237(LCDR1), SEQ ID NO 238(LCDR2) and SEQ ID NO 239(LCDR 3); (III) SEQ ID NO:370(HCDR1), SEQ ID NO:371(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3); or (IV) SEQ ID NO:372(HCDR1), SEQ ID NO:373(HCDR2), SEQ ID NO:232(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2) and SEQ ID NO:239(LCDR 3). In some embodiments, ZZ12_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:367(HCDR1), SEQ ID NO:368(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3). In some embodiments, ZZ12_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:369(HCDR1), SEQ ID NO:368(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3). In some embodiments, ZZ12_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:370(HCDR1), SEQ ID NO:371(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3). In some embodiments, ZZ12_ LALA may be defined as comprising or having the amino acid sequence: 372(HCDR1), 373(HCDR2), 232(HCDR3), 243(LCDR1), 241(LCDR2), and 239(LCDR 3). In some embodiments, ZZ12_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 374 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 244. In some embodiments, ZZ12_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 376 and a light chain comprising or having the amino acid sequence of SEQ ID No. 246.

For example, ZZ13_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 378(HCDR1), 379(HCDR2), 228(HCDR3), 237(LCDR1), 238(LCDR2) and 239(LCDR 3); (II) SEQ ID NO:380(HCDR1), SEQ ID NO:379(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2) and SEQ ID NO:239(LCDR 3); (III) SEQ ID NO:381(HCDR1), SEQ ID NO:382(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3); or (IV) SEQ ID NO 383(HCDR1), SEQ ID NO 384(HCDR2), SEQ ID NO 232(HCDR3), SEQ ID NO 243(LCDR1), SEQ ID NO 241(LCDR2), and SEQ ID NO 239(LCDR 3). In some embodiments, ZZ13_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:378(HCDR1), SEQ ID NO:379(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3). In some embodiments, ZZ13_ LALA may be defined as comprising or having the amino acid sequence: 380(HCDR1) SEQ ID NO:379(HCDR2), 228(HCDR3) SEQ ID NO:237(LCDR1), 238(LCDR2) and 239(LCDR 3). In some embodiments, ZZ13_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:381(HCDR1), SEQ ID NO:382(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3). In some embodiments, ZZ13_ LALA may be defined as comprising or having the amino acid sequence: (IV) SEQ ID NO:383(HCDR1), SEQ ID NO:384(HCDR2), SEQ ID NO:232(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:239(LCDR 3). In some embodiments, ZZ13_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 385 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 244. In some embodiments, ZZ13_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 387 and a light chain comprising or having the amino acid sequence of SEQ ID No. 246.

For example, ZZ14_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 270(HCDR1), 389(HCDR2), 331(HCDR3), 337(LCDR1), 338(LCDR2) and 339(LCDR3) SEQ ID NO; (II) SEQ ID NO:273(HCDR1), SEQ ID NO:389(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:390(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 275(HCDR1), SEQ ID NO 391(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3). In some embodiments, ZZ14_ LALA may be defined as comprising or having the amino acid sequence: 270(HCDR1), 389(HCDR2), 331(HCDR3), 337(LCDR1), 338(LCDR2), and 339(LCDR3) SEQ ID NO. In some embodiments, ZZ14_ LALA may be defined as comprising or having the amino acid sequence: 273(HCDR1) SEQ ID NO:389(HCDR2) SEQ ID NO:331(HCDR3), 337(LCDR1) SEQ ID NO:338(LCDR2) and 339(LCDR3) SEQ ID NO. In some embodiments, ZZ14_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:32(HCDR1), SEQ ID NO:390(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3). In some embodiments, ZZ14_ LALA may be defined as comprising or having the amino acid sequence: 275(HCDR1), 391(HCDR2), 332(HCDR3), 343(LCDR1), 341(LCDR2), and 339(LCDR3) SEQ ID NO. In some embodiments, ZZ14_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:392 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 344. In some embodiments, ZZ14_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 394 and a light chain comprising or having the amino acid sequence of SEQ ID No. 346.

For example, ZZ15_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 396(HCDR1), 397(HCDR2), 331(HCDR3), 337(LCDR1), 338(LCDR2) and 339(LCDR3) SEQ ID; (II) SEQ ID NO:398(HCDR1), SEQ ID NO:397(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:399(HCDR1), SEQ ID NO:400(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 401(HCDR1), SEQ ID NO 402(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3). In some embodiments, ZZ15_ LALA may be defined as comprising or having the amino acid sequence: 396(HCDR1), 397(HCDR2), 331(HCDR3), 337(LCDR1), 338(LCDR2), and 339(LCDR 3). In some embodiments, ZZ15_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:398(HCDR1), SEQ ID NO:397(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2), and SEQ ID NO:339(LCDR 3). In some embodiments, ZZ15_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:399(HCDR1), SEQ ID NO:400(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3). In some embodiments, ZZ15_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO 401(HCDR1), SEQ ID NO 402(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2), and SEQ ID NO 339(LCDR 3). In some embodiments, ZZ15_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:403 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 344. In some embodiments, ZZ15_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 405 and a light chain comprising or having the amino acid sequence of SEQ ID No. 346.

For example, ZZ16_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 407(HCDR1), 408(HCDR2), 331(HCDR3), 337(LCDR1), 338(LCDR2) and 339(LCDR3) SEQ ID NO; (II) SEQ ID NO:409(HCDR1), SEQ ID NO:408(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:410(HCDR1), SEQ ID NO:411(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 412(HCDR1), SEQ ID NO 413(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3). In some embodiments, ZZ16_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:407(HCDR1), SEQ ID NO:408(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2), and SEQ ID NO:339(LCDR 3). In some embodiments, ZZ16_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:409(HCDR1), SEQ ID NO:408(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2), and SEQ ID NO:339(LCDR 3). In some embodiments, ZZ16_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:410(HCDR1), SEQ ID NO:411(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3). In some embodiments, ZZ16_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:412(HCDR1), SEQ ID NO:413(HCDR2), SEQ ID NO:332(HCDR3), SEQ ID NO:343(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:339(LCDR 3). In some embodiments, ZZ16_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 414 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 344. In some embodiments, ZZ16_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 416 and a light chain comprising or having the amino acid sequence of SEQ ID No. 346.

For example, ZZ17_ LALA may be defined as having three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from: (I) 418(HCDR1), 419(HCDR2), 331(HCDR3), 337(LCDR1), 338(LCDR2) and 339(LCDR3) SEQ ID; (II) SEQ ID NO:420(HCDR1), SEQ ID NO:419(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:421(HCDR1), SEQ ID NO:422(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO:423(HCDR1), SEQ ID NO:424(HCDR2), SEQ ID NO:332(HCDR3), SEQ ID NO:343(LCDR1), SEQ ID NO:341(LCDR2) and SEQ ID NO:339(LCDR 3). In some embodiments, ZZ17_ LALA may be defined as comprising or having the amino acid sequence: 418(HCDR1) SEQ ID NO:419(HCDR2) SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2), and SEQ ID NO:339(LCDR 3). In some embodiments, ZZ17_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:420(HCDR1), SEQ ID NO:419(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2), and SEQ ID NO:339(LCDR 3). In some embodiments, ZZ17_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:421(HCDR1), SEQ ID NO:422(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3). In some embodiments, ZZ17_ LALA may be defined as comprising or having the amino acid sequence: SEQ ID NO:423(HCDR1), SEQ ID NO:424(HCDR2), SEQ ID NO:332(HCDR3), SEQ ID NO:343(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:339(LCDR 3). In some embodiments, ZZ17_ LALA may be defined as comprising or having a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:425 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO: 344. In some embodiments, ZZ17_ LALA may be defined as comprising or having a heavy chain comprising or having the amino acid sequence of SEQ ID No. 427 and a light chain comprising or having the amino acid sequence of SEQ ID No. 346.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 13 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 13 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 24 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 24. In some other embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 15 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 15 and a light chain comprising or having the amino acid sequence of SEQ ID No. 26 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 26.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 37 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 48 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 48. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 39 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 39 and a light chain comprising or having the amino acid sequence of SEQ ID No. 50 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 50.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 61 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 61 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 72 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 72. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 63 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 63 and a light chain comprising or having the amino acid sequence of SEQ ID No. 74 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 74.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:85 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:85 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:96 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 96. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 87 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 87 and a light chain comprising or having the amino acid sequence of SEQ ID No. 98 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 98.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 103 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 103 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 24 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 24. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 105 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 105 and a light chain comprising or having the amino acid sequence of SEQ ID No. 26 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 26.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 103 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 103 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 24 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 24. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 108 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 108 and a light chain comprising or having the amino acid sequence of SEQ ID No. 26 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 26.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 115 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 115 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 24 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 24. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 117 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 117 and a light chain comprising or having the amino acid sequence of SEQ ID No. 26 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 26.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 122 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 48 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 48. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 124 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 124 and a light chain comprising or having the amino acid sequence of SEQ ID No. 50 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 50.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 37 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 128 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 128. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 39 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 39 and a light chain comprising or having the amino acid sequence of SEQ ID No. 130 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 130.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 37 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 136 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 136. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 39 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 39 and a light chain comprising or having the amino acid sequence of SEQ ID No. 138 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 138.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 37 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 136 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 136. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID NO 141 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 141 and a light chain comprising or having the amino acid sequence of SEQ ID NO 138 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 138.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 37 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 147 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 147. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 39 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 39 and a light chain comprising or having the amino acid sequence of SEQ ID No. 149 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 149.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:154 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:154 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:24 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 24. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 156 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 156 and a light chain comprising or having the amino acid sequence of SEQ ID No. 26 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 26.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:154 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:154 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:24 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 24. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 159 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 159 and a light chain comprising or having the amino acid sequence of SEQ ID No. 26 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 26.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 161 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 161 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 24 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 24. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 163, or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 163, and a light chain comprising or having the amino acid sequence of SEQ ID No. 26, or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 26.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:168 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:168 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:24 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 24. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 170 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 170 and a light chain comprising or having the amino acid sequence of SEQ ID No. 26 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 26.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO 37 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO 174 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 174. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 39 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 39 and a light chain comprising or having the amino acid sequence of SEQ ID No. 176 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 176.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 37 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 180 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 180. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 39 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 39 and a light chain comprising or having the amino acid sequence of SEQ ID No. 182 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 182.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 37 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 37 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 186 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 186. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 39 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 39 and a light chain comprising or having the amino acid sequence of SEQ ID No. 188 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 188.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 193 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 193 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 136 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 136. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 195, or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 195, and a light chain comprising or having the amino acid sequence of SEQ ID No. 138, or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 138.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO 193 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 193 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO 174 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 174. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 195, or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 195, and a light chain comprising or having the amino acid sequence of SEQ ID No. 176, or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 176.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO 201 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO 128 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 128. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 203 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 203 and a light chain comprising or having the amino acid sequence of SEQ ID No. 130 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 130.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 122 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 128 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 128. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 208 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 208 and a light chain comprising or having the amino acid sequence of SEQ ID No. 130 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 130.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:201 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:136 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 136. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 203 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 203 and a light chain comprising or having the amino acid sequence of SEQ ID No. 138 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 138.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 122 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 136 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 136. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 208 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 208 and a light chain comprising or having the amino acid sequence of SEQ ID No. 138 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 138.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO 201 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO 147 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 147. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 203 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 203 and a light chain comprising or having the amino acid sequence of SEQ ID No. 149 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 149.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 122 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 147 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 147. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 208 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 208 and a light chain comprising or having the amino acid sequence of SEQ ID No. 149 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 149.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO 201 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO 174 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 174. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 203 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 203 and a light chain comprising or having the amino acid sequence of SEQ ID No. 176 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 176.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 122 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 174 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 174. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 208 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 208 and a light chain comprising or having the amino acid sequence of SEQ ID No. 176 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 176.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:201 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:180 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 180. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 203 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 203 and a light chain comprising or having the amino acid sequence of SEQ ID No. 182 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 182.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 122 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 180 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 180. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 208 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 208 and a light chain comprising or having the amino acid sequence of SEQ ID No. 182 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 182.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO 201 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 201 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO 186 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 186. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 203 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 203 and a light chain comprising or having the amino acid sequence of SEQ ID No. 188 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 188.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 122 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 122 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 186 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 186. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 208 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 208 and a light chain comprising or having the amino acid sequence of SEQ ID No. 188 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 188.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 233 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 233 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 244 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 244. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 235 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 235 and a light chain comprising or having the amino acid sequence of SEQ ID No. 246 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 246.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:256 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:256 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:266 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 266. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 258 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 258 and a light chain comprising or having the amino acid sequence of SEQ ID No. 268 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 268.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 278 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 278 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 287 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 287. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 280 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 280 and a light chain comprising or having the amino acid sequence of SEQ ID No. 289 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 289.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 300 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 300 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 306 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 306. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 302 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 302 and a light chain comprising or having the amino acid sequence of SEQ ID No. 308 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 308.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:316 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:316 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:327 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 327. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 318 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 318 and a light chain comprising or having the amino acid sequence of SEQ ID No. 329 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 329.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 333 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 333 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 344 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 344. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 335 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 335 and a light chain comprising or having the amino acid sequence of SEQ ID No. 346 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 346.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:350 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:350 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:357 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 357. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID NO 352 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 352 and a light chain comprising or having the amino acid sequence of SEQ ID NO 359 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO 359.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:350 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:350 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:363 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 363. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 352 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 352 and a light chain comprising or having the amino acid sequence of SEQ ID No. 365 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 365.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 374 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 374 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 244 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 244. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 376 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 376 and a light chain comprising or having the amino acid sequence of SEQ ID No. 246 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 246.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 385 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 385 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 244 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 244. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 387 or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 387 and a light chain comprising or having the amino acid sequence of SEQ ID No. 246 or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 246.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:392 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:392 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:344 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 344. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 394 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 394 and a light chain comprising or having the amino acid sequence of SEQ ID No. 346 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 346.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:403 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:403 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:344 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 344. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 405 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 405 and a light chain comprising or having the amino acid sequence of SEQ ID No. 346 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 346.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID No. 414 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 414 and a light chain variable region comprising or having the amino acid sequence of SEQ ID No. 344 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 344. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 416 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 416 and a light chain comprising or having the amino acid sequence of SEQ ID No. 346 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID No. 346.

In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain variable region comprising or having the amino acid sequence of SEQ ID NO:425 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO:425 and a light chain variable region comprising or having the amino acid sequence of SEQ ID NO:344 or a sequence having at least 95%, 96%, 97%, 98% or 99% identity to SEQ ID NO: 344. In some embodiments, the antibody or antigen-binding fragment comprises a heavy chain comprising or having the amino acid sequence of SEQ ID No. 427, or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 427, and a light chain comprising or having the amino acid sequence of SEQ ID No. 346, or a sequence having at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID No. 346.

Sets of exemplary anti-NPR 1 antibodies of the present disclosure are listed in Table 3 or Table 4 in terms of CDRs (i.e., the numbers in these tables represent sequence identifiers, thus "28" represents "SEQ ID NO: 28") or amino acid consensus sequences. Where a plurality of values are shown in Table 3, they can be used for the CDR's alternatives (i.e., where SEQ ID Nos: 29, 119 and 190 are listed as HCDR2, they can be used for the CDR's alternatives). In Table 4, those amino acids enclosed in parentheses and separated by slashes indicate the substituted amino acid at that position (e.g., "(A/V)" indicates the position where the amino acid may be alanine or valine). In some embodiments, the antibody has the heavy and light chain CDRs of any of the antibodies described in table 3 or table 4. In some embodiments, the anti-NPR 1 antibody is a four chain antibody (also referred to as a whole antibody) comprising two heavy chains and two light chains. In some embodiments, the anti-NPR 1 antibody is an antigen-binding fragment of an intact antibody, e.g., a functional fragment of an intact antibody selected from those listed in table 3 or table 4, which retains the ability to bind NPR1 and/or provide intact antibody function (e.g., activate NPR1 in the absence of ANP).

TABLE 3 exemplary set of anti-NPR 1 antibodies based on CDRs

TABLE 4 exemplary set of anti-NPR 1 antibodies based on consensus sequences

In some embodiments, an antibody or antigen-binding fragment thereof as provided herein is conjugated to (a) human NPR 1; and (b) mouse NPR1 and/or rat NPR 1.

In some embodiments, an antibody or antigen-binding fragment thereof as provided herein is conjugated to (a) human NPR 1; and (b) cynomolgus monkey NPR1 binding. In some embodiments, the antibody or antigen-binding fragment thereof is therapeutic. As defined herein, a therapeutic antibody is an effective and stable antibody.

Antibodies that bind to the same epitope as the anti-NPR 1 antibodies of the disclosure

In another embodiment, the disclosure provides an antibody or antigen-binding fragment thereof that binds to the same epitope as one or more anti-NPR 1 antibodies (e.g., WW06) described herein. Such antibodies:

(i) binding to NPR 1;

(ii) is an agonist of NPR 1;

(iii) is ANP competitive; and

binds to the same epitope in NPR1 as antibody WW 06.

In another embodiment, the disclosure provides an antibody or antigen-binding fragment thereof that binds to the same epitope as one or more anti-NPR 1 antibodies (e.g., XX16) described herein. Such antibodies:

(i) binding to NPR 1;

(ii) Is an agonist of NPR 1;

(iii) is ANP non-competitive; and

binds to the same epitope in NPR1 as XX 16.

In another embodiment, the disclosure provides an antibody or antigen-binding fragment thereof that binds to the same epitope as one or more anti-NPR 1 antibodies (e.g., WW03) described herein. Such antibodies:

(i) binding to NPR 1;

(ii) is an agonist of NPR 1;

(iii) is ANP non-competitive; and

binds to the same epitope in NPR1 as WW 03.

After crystallization and structure determination, the binding regions of preferred antibodies of the present disclosure have been more clearly defined. Such binding is defined herein to include covalent and non-covalent bonds.

Thus, the present disclosure provides ANP competitive antibodies that bind the same epitope as WW 06. In some embodiments, the disclosure provides antibodies that bind to an epitope of human NPR1 protein (accession number P16066; SEQ ID NO:1) comprising amino acids 188, 192, 194, 197, 201, 208, and 219. In some embodiments, the disclosure provides antibodies that bind to an epitope of human NPR1 protein (accession number P16066; SEQ ID NO:1) comprising amino acids 188, 192, 194, 197, 201, 208, 219, and 295. In some embodiments, the disclosure provides antibodies that bind to an epitope within amino acid numbers 188-198 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to at least 2, 3, or 4 amino acid residues within amino acid numbers 188-198 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to an epitope within amino acid numbers 201-208 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to at least 2 amino acids within amino acid numbers 201-208 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to at least 2, 3, or 4 amino acid residues within amino acid numbers 188-198 of SEQ ID NO:1 and bind to at least 2 amino acids within amino acid numbers 201-208 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to an epitope comprising at least one amino acid residue in each of (i) amino acids 188-198 of SEQ ID NO:1, (ii) amino acids 201-208 of SEQ ID NO:1, (iii) amino acids 215-238 of SEQ ID NO:1, and (iv) amino acids 294-297 of SEQ ID NO: 1.

The present disclosure provides ANP non-competitive antibodies that bind the same epitope as WW 03. In some embodiments, the disclosure provides antibodies that bind to an epitope of human NPR1 protein (accession number P16066; SEQ ID NO:1) comprising

amino acids

82, 102, 103, 105, 106, 109, 132, and 375. In some embodiments, the disclosure provides antibodies that bind to an epitope of human NPR1 protein (accession number P16066; SEQ ID NO:1) comprising

amino acids

79, 82, 99, 102, 103, 105, 106, 109, 131, 132, and 375. In some embodiments, the disclosure provides antibodies that bind to an epitope within amino acid numbers 99-111 of SEQ ID NO. 1. In some embodiments, the disclosure provides antibodies that bind to at least 2, 3, 4, 5, or 6 amino acids within amino acid numbers 99-111 of SEQ ID No. 1. In some embodiments, the disclosure provides antibodies that bind to at least 2, 3, 4, 5, 6, 7, or 8 amino acid residues within amino acid numbers 99-133 of SEQ ID No. 1. In some embodiments, the disclosure provides antibodies that bind to an epitope within amino acid numbers 131 and 134 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to at least 2 amino acids within amino acid numbers 131 and 134 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to at least 2, 3, 4, 5, or 6 amino acids within amino acid numbers 99-111 of SEQ ID NO:1 and bind to at least 2 amino acids within amino acid numbers 131 and 134 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to an epitope comprising at least one amino acid residue in each of (i) amino acids 99-111 of SEQ ID NO:1, (ii) amino acids 131-134 of SEQ ID NO:1, and (iii) amino acids 374-375 of SEQ ID NO: 1. Optionally, the antibody may additionally bind to amino acids 79 and/or 82 of SEQ ID NO. 1.

The present disclosure additionally provides ANP non-competitive antibodies that bind the same epitope as XX 16. In some embodiments, the disclosure provides antibodies that bind to an epitope of human NPR1 protein (accession number P16066; SEQ ID NO:1) comprising

amino acids

82, 102, 103, 105, 106, 109, 132, and 375. In some embodiments, the disclosure provides antibodies that bind to an epitope of the human NPR1 protein (accession number P16066; SEQ ID NO:1) comprising

amino acids

34, 82, 102, 103, 105, 106, 107, 109, 132, 133, 375, and 378. In some embodiments, the disclosure provides antibodies that bind to an epitope within amino acid numbers 102-111 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to at least 2, 3, 4, 5, or 6 amino acids within amino acid numbers 102-111 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to an epitope within amino acid numbers 131 and 134 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to at least 2 amino acids within amino acid numbers 131 and 134 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to at least 2, 3, 4, 5, or 6 amino acids within amino acid numbers 102-111 of SEQ ID NO:1 and bind to at least 2 amino acids within amino acid numbers 131-134 of SEQ ID NO: 1. In some embodiments, the disclosure provides antibodies that bind to an epitope comprising at least one amino acid residue in each of (i) amino acids 102-111 of SEQ ID NO:1, (ii) amino acids 131-134 of SEQ ID NO:1, and (iii) amino acids 374-378 of SEQ ID NO: 1. Optionally, the antibody may additionally bind to amino acids 34, 76 and/or 82 of SEQ ID NO. 1.

Additional antibodies can be identified based on their ability to cross-compete (e.g., competitively inhibit binding of other antibodies of the disclosure in a statistically significant manner) with other antibodies of the disclosure (e.g., XX16, WW06, or WW03) in a standard NPR1 binding assay. The ability of the test antibody to inhibit the binding of an antibody of the present disclosure to human NPR1 demonstrates that the test antibody can compete with the antibody for binding to human NPR 1; according to a non-limiting theory, such an antibody may bind to the same or related (e.g., structurally similar or spatially close) epitope on human NPR1 as the antibody it competes for. In a certain embodiment, an antibody that binds to the same epitope on human NPR1 as an antibody of the present disclosure is a human antibody (e.g., a human monoclonal antibody or antigen-binding fragment thereof). Such antibodies can be prepared and isolated as described herein.

Engineered and modified antibodies

Antibodies of the present disclosure can be prepared using antibodies having one or more of the VH and/or VL sequences set forth herein as starting materials to engineer modified antibodies that may have altered properties compared to the starting antibody. Antibodies can be engineered by modifying one or more residues within one or both variable regions (i.e., VH and/or VL), e.g., within one or more CDR regions and/or within one or more framework regions. Additionally or alternatively, the antibody may be engineered by modifying residues within one or more constant regions, for example to alter one or more effector functions of the antibody.

One type of variable region engineering that can be performed is antibody binding region/paratope or CDR grafting. Since the complement sequence is responsible for most antibody-antigen interactions, recombinant antibodies that mimic the properties of a particular naturally occurring antibody can be expressed by constructing expression vectors that include the CDR/complement sequences from a particular naturally occurring antibody grafted onto framework sequences from different antibodies with different properties (see, e.g., Riechmann, L. et al, 1998Nature [ Nature ]332: 323-327; Jones, P. et al, 1986Nature [ Nature ]321: 522-525; Queen, C. et al, 1989Proc. Natl. Acad. Sci. U.S. A. [ Proc. Natl. Acad. Sci. U.S. 20; U.S. Pat. 10086: 10029-089; U.S. Pat. No. 5,225,539, and U.S. Pat. Nos. 5,530,101, 5,585,089, 5,693,762, and 6,180,370; the contents of each of which are hereby incorporated by reference).

Accordingly, another embodiment of the disclosure relates to an isolated anti-NPR 1 antibody, or antigen-binding fragment thereof, comprising an antigen-binding portion thereof comprising a heavy chain variable region comprising the CDR sequences of an antibody or set of antibodies as set forth in table 2, table 3, or table 4. Thus, such antibodies contain the V of a monoclonal antibody _HAnd V_LThe CDR sequences may also contain different framework sequences from these antibodies.

Such framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences. For example, germline DNA Sequences for Human heavy and light chain variable region genes can be found in the "VBase" Human germline sequence database (available from the Internet www [ dot ] mrc-cpe [ dot ] cam [ dot ] ac [ dot ] uk/VBase), and in kabat, E.A., et al, 1991Sequences of Proteins of Immunological Interest [ immunologically related protein Sequences ], fifth edition, U.S. department of Health and public service (U.S. Department of Health and Human Services), NIH publication Nos. 91-3242; tomlinson, i.m. et al, 1992j.fol.biol. [ journal of molecular biology ]227: 776-; and Cox, J.P.L. et al, 1994Eur.J Immunol. [ J. Eur. Immunol ]24: 827-; the contents of each are hereby incorporated by reference.

Examples of framework sequences for use in antibodies or antigen-binding fragments of the disclosure are those that are structurally similar to the framework sequences used by selected antibodies of the disclosure, e.g., consensus sequences and/or framework sequences used by antibodies or antigen-binding fragments of the disclosure. Can be combined with V _HCDR1, 2 and 3 sequences and V_LThe CDR1, 2 and 3 sequences can be grafted onto framework regions having the same sequence as the framework regions found in the germline immunoglobulin gene from which the framework sequences are derived, or the CDR sequences can be grafted onto framework regions containing one or more mutations as compared to the germline sequence. For example, it has been found beneficial in certain circumstances to mutate residues within the framework regions to maintain or enhance the antigen binding ability of an antibody (see, e.g., U.S. Pat. nos. 5,530,101, 5,585)089, 5,693,762 and 6,180,370; the contents of each of which are hereby incorporated by reference).

Another type of variable region modification is to modify V_HAnd/or V_LAmino acid residues within the CDR1, CDR2, and/or CDR3 regions are mutated to improve one or more binding properties (e.g., affinity) of the antibody of interest, referred to as "affinity maturation. Site-directed mutagenesis or PCR-mediated mutagenesis can be performed to introduce one or more mutations, and the effect on antibody binding or other functional property of interest can be assessed in an in vitro or in vivo assay as described herein. Conservative modifications (as discussed above) may also be introduced. The mutation may be an amino acid substitution, addition or deletion. Furthermore, typically no more than one, two, three, four or five residues within a CDR region are altered.

Transplantation of antigen binding domains into alternative frameworks or scaffolds

A variety of antibody/immunoglobulin frameworks or scaffolds may be used, so long as the resulting polypeptide comprises at least one binding region that specifically binds NPR 1. Such frameworks or scaffolds include human immunoglobulins or fragments thereof of 5 major idiotypes (such as those disclosed elsewhere herein), and immunoglobulins of other animal species, preferably with humanization aspects. In this regard, single heavy chain antibodies (such as those identified in camelids) are of particular interest. Those skilled in the art continue to discover and develop novel frameworks, scaffolds and fragments.

In one aspect, the disclosure relates to the generation of non-immunoglobulin based antibodies using non-immunoglobulin scaffolds on which CDRs of the disclosure can be grafted. Known or future non-immunoglobulin frameworks and scaffolds may be used as long as they comprise a binding region specific for NPR1, e.g., as disclosed for the antibodies described herein, including but not limited to XX16, WW03 or WW 06. Such compounds are referred to herein as "polypeptides comprising a target-specific binding region". Examples of non-immunoglobulin frameworks are further described in the following sections (camelid antibodies and non-antibody scaffolds).

Camelidae animal antibodies

Antibody proteins obtained from members of the camel and dromedary (bactrian and Calelus demanderius) families, including new members of the world such as llama species (Lama paccos, llama and leptospora) have been characterized with respect to size, structural complexity and antigenicity in human subjects. Certain IgG antibodies from this mammalian family, as found in nature, lack a light chain and are therefore structurally different from the typical four-chain quaternary structure with two heavy chains and two light chains of antibodies from other animals, see WO 94/04678, the contents of which are hereby incorporated by reference.

Regions of camelid antibodies, which are small single variable domains identified as VHHs, are obtained by genetic engineering to produce small proteins with high affinity for the target, resulting in low molecular weight antibody-derived proteins known as "camelid nanobodies". See US5,759,808; see also Stijlemans, B, et al, 2004J Biol Chem [ J. Biol. Chem ]279: 1256-S.sub.1261; dumoulin, M. et al, 2003Nature [ Nature ]424: 783-; pleschberger, M. et al, 2003Bioconjugate Chem Bioconjugate chemistry 14: 440-; cortex-Retamozo, V. et al, 2002Int J Cancer [ J. International Cancer ]89: 456-62; and Lauwereys, M. et al, 1998EMBO J [ J. European journal of molecular biology ]17: 3512-3520; the contents of each are hereby incorporated by reference. Engineered libraries of camelid antibodies and antibody fragments are commercially available, for example, from ebolingx, outlex, rhizot, belgium. Like other antibodies of non-human origin, the amino acid sequence of a camelid antibody can be recombinantly altered to obtain a sequence more closely resembling a human sequence, i.e., the nanobody can be "humanized". Thus, the natural low antigenicity of camelid antibodies to humans can be further reduced.

The molecular weight of camelid nanobodies is about one tenth of that of human IgG molecules, and the physical diameter of the protein is only a few nanometers. One consequence of the small size is the ability of camelid nanobodies to bind to antigenic sites that are functionally invisible to larger antibody proteins, i.e., camelid nanobodies may be used as reagents to detect antigens that are cryptic to the use of classical immunological techniques, as well as possible therapeutic agents. Thus, yet another consequence of the small size is that camelid nanobodies may be inhibited due to binding to specific sites in the groove or slit of the target protein and thus may have the ability to function more closely like classical low molecular weight drugs compared to classical antibodies.

The low molecular weight and compact size also result in camelid nanobodies that are extremely thermostable, stable to extreme pH and proteolytic digestion, and poorly antigenic. Another consequence is that camelid nanobodies move easily from the circulatory system into tissues, even across the blood-brain barrier and can treat disorders affecting nervous tissues. Nanobodies may further facilitate transport of drugs across the blood-brain barrier, see US 2004/0161738, the contents of which are hereby incorporated by reference. These features combined with low antigenicity in humans show great therapeutic potential. Furthermore, these molecules can be fully expressed in prokaryotic cells such as E.coli and can be expressed as fusion proteins with phage.

Accordingly, the disclosure features camelid antibodies or nanobodies with high affinity for NPR 1. In one embodiment, a camelid antibody or nanobody is obtained by grafting CDR sequences of the heavy or light chain of a human antibody of the present disclosure into a nanobody or single domain antibody framework sequence, as described, for example, in WO 94/04678 (the contents of which are hereby incorporated by reference).

Framework or Fc engineering

Engineered antibodies of the disclosure include those that: wherein framework residues within the VH and/or VL have been modified, for example to improve one or more properties of the antibody. Typically, such framework modifications are made to reduce the immunogenicity of the antibody. Antibodies of the present disclosure may be modified in one or more ways, including each of the ways described herein.

For example, one approach is to "back mutate" one or more framework residues to the corresponding germline sequence. Rather, an antibody that has undergone somatic mutation may contain framework residues that differ from the germline sequence from which the antibody was derived. Such residues can be identified by comparing the antibody framework sequences to the germline sequences of the derivative antibody. In order to restore the framework region sequences to their germline configuration, somatic mutations can be "back-mutated" to germline sequences by, for example, site-directed mutagenesis or PCR-mediated mutagenesis. Such "back-mutated" antibodies and additional modifications described herein are also intended to be encompassed by the present disclosure.

Another type of framework modification includes mutating one or more residues within the framework regions or even within one or more CDR regions to remove T cell epitopes, thereby reducing the potential immunogenicity of the antibody. This method is also referred to as "deimmunization" and is described in more detail in US 2003/0153043, the content of which is hereby incorporated by reference.

In addition to or as an alternative to modifications made within the framework or CDR regions, antibodies of the disclosure can be engineered to include modifications within the Fc region, typically in order to alter one or more functional properties of the antibody, such as serum half-life, complement binding, Fc receptor binding, and/or antigen-dependent cellular cytotoxicity. Furthermore, an antibody of the present disclosure can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or modified to alter its glycosylation, thereby again altering one or more functional properties of the antibody. Each of these embodiments is described in more detail below. The numbering of residues in the Fc region is that of the EU index of kabat.

In one embodiment, the hinge region of CH1 is modified such that the number of cysteine residues in the hinge region is altered, e.g., increased or decreased. The process is further described in US5,677,425, the contents of which are hereby incorporated by reference. The number of cysteine residues in the CH1 hinge region is altered, for example, to facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody.

In another embodiment, the Fc hinge region of the antibody is mutated to shorten the biological half-life of the antibody. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of the Fc-hinge fragment such that the antibody has impaired staphylococcal protein a (SpA) binding relative to native Fc-hinge domain SpA binding. This method is described in more detail in US 6,165,745, the contents of which are hereby incorporated by reference.

In another embodiment, the antibody is modified to increase its biological half-life. Various methods may be employed. For example, one or more of the following mutations may be introduced: T252L, T254S, T256F, as described in US 6,277,375, the contents of which are hereby incorporated by reference. Alternatively, to increase biological half-life, antibodies may be altered within the CH1 or CL region to contain salvage receptor binding epitopes taken from the two loops of the CH2 domain in the Fc region of IgG, as described in US 5,869,046 and US 6,121,022, the respective contents of which are hereby incorporated by reference.

In yet other embodiments, the Fc region is altered by substituting at least one amino acid residue with a different amino acid residue to alter the effector function of the antibody. For example, one or more amino acids may be substituted with different amino acid residues such that the antibody has an altered affinity for the effector ligand, but retains the antigen binding ability of the parent antibody. The affinity-altering effector ligand may be, for example, an Fc receptor or the C1 component of complement. The process is described in more detail in US 5,624,821 and US 5,648,260, the contents of each of which are hereby incorporated by reference.

To minimize ADCC activity of the antibody, specific mutations within the Fc region produce "Fc-silent" antibodies with minimal interaction with effector cells. In general, "IgG Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. The human IgG heavy chain Fc region is generally defined as comprising the amino acid residues from position C226 or from position P230 to the carboxy-terminus of an IgG antibody. The numbering of residues in the Fc region is that of the EU index of kabat. For example, the C-terminal lysine (residue K447) of the Fc region may be removed during production or purification of the antibody.

Silent effector functions can be obtained by mutation of the Fc region of an antibody. See, e.g., LALA and N297A (Strohl, W.,2009, Curr. Opin. Biotechnol. [ Current Biotechnology View ] Vol. 20(6) 685.: 685-; and D265A (Baudino et al, 2008, j. immunol. [ journal of immunology ]181:6664-69) see also Heusser et al, WO 2012065950; the contents of each are hereby incorporated by reference. In particular, residues 234 and/or 235 may be mutated, optionally to alanine. Thus, in one embodiment, an antibody according to the present disclosure has a mutation in the Fc region at one or both of amino acids 234 and 235. This substitution of both amino acids 234 and 235 results in a decrease in ADCC activity. An example of such a mutation is a LALA mutant comprising L234A and L235A mutations in the IgGl Fc amino acid sequence. Another example of a silent IgGl antibody is a DAPA (D265A, P329A) mutation (US 6,737,056, the content of which is hereby incorporated by reference). Another silent IgGl antibody comprises a N297A mutation, which results in an aglycosylated/aglycosylated antibody. Fc-silenced antibodies produce no or low ADCC activity, meaning that the Fc-silenced antibody exhibits less than 50% of the specific cell lysis for ADCC activity. No ADCC activity means that the Fc-silencing antibody exhibits less than 1% ADCC activity (specific cell lysis).

In another example, one or more amino acids selected from the group consisting of amino acid residues may be substituted with a different amino acid residue such that the antibody has altered C1q binding and/or reduced or eliminated Complement Dependent Cytotoxicity (CDC). This process is described in more detail in US6,194,551, the contents of which are hereby incorporated by reference.

In another embodiment, one or more amino acid residues are altered, thereby altering the ability of the antibody to fix complement. This process is further described in WO 94/29351, the content of which is hereby incorporated by reference.

In another embodiment, the Fc region is modified to increase the ability of the antibody to mediate antibody-dependent cellular cytotoxicity (ADCC) and/or to increase the affinity of the antibody for the fey receptor by modifying one or more amino acids. This process is further described in WO 00/42072, the content of which is hereby incorporated by reference. Furthermore, the binding sites for Fc γ Rl, Fc γ RII, Fc γ RIII and FcRn on human IgG1 have been mapped and variants with improved binding have been described (see Shields, r.l. et al, 2001j.biol.chen. [ journal of biochemistry ]276: 6591-.

In yet another embodiment, the glycosylation of the antibody is modified. For example, non-glycosylated antibodies (i.e., antibodies lacking glycosylation) can be prepared. Glycosylation can be altered, for example, to increase the affinity of an antibody for an "antigen". Such carbohydrate modifications can be achieved, for example, by altering one or more glycosylation sites within the antibody sequence. For example, one or more amino acid substitutions can be made that eliminate one or more variable region framework glycosylation sites, thereby eliminating glycosylation at such sites. Such aglycosylation may increase the affinity of the antibody for the antigen. Such methods are described in more detail in US 5,714,350 and US 6,350,861, the contents of each of which are hereby incorporated by reference.

Additionally or alternatively, antibodies with altered glycosylation patterns can be made, such as low fucosylated antibodies with reduced amounts of fucosyl residues or antibodies with increased bisecting GlcNac structures. Such altered glycosylation patterns have been shown to increase the ADCC capacity of the antibody. Such carbohydrate modification can be achieved, for example, by expressing the antibody in a host cell with an altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and can be used as host cells in which to express recombinant antibodies of the disclosure, thereby producing antibodies with altered glycosylation. For example, EP 1,176,195 (the content of which is hereby incorporated by reference) describes a cell line with a functionally disrupted FUT8 gene encoding a fucosyltransferase such that antibodies expressed in such cell line exhibit low fucosylation. WO 03/035835 describes a variant CHO cell line Lecl3 cell which has a reduced ability to attach fucose to Asn (297) linked carbohydrates, and also results in low fucosylation of antibodies expressed in the host cell (see also Shields, r.l. et al, 2002j. biol. chem. [ J. biochem ]277: 26733. 26740). WO 99/54342 describes cell lines engineered to express glycoprotein-modified glycosyltransferases (e.g.,. beta. (1,4) -N-acetylglucosaminyltransferase III (GnTIII)) such that antibodies expressed in the engineered cell lines exhibit an increased bisecting GlcNac structure that results in increased ADCC activity of the antibody (see also Umana et al, nat 1999. Biotech. [ Nature Biotechnology ]17: 176-180). The contents of each of the foregoing applications and references are hereby incorporated by reference.

Another modification of the antibodies herein contemplated by the present disclosure is pegylation. Antibodies can be pegylated, for example, to increase the biological (e.g., serum) half-life of the antibody. To pegylate an antibody, the antibody or fragment thereof is typically reacted with polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups are attached to the antibody or antibody fragment. Pegylation can be performed by acylation or alkylation reactions using reactive PEG molecules (or similar reactive water-soluble polymers). As used herein, the term "polyethylene glycol" is intended to encompass any form of PEG that has been used to derivatize other proteins, such as mono (C1-C10) alkoxy-or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certain embodiments, the antibody to be pegylated is an aglycosylated antibody. Methods of pegylating proteins are known in the art and can be applied to the antibodies of the present disclosure. See, e.g., EP 0154316 and EP 0401384, the respective contents of which are hereby incorporated by reference.

Another modification of an antibody contemplated by the present disclosure is a conjugate or protein fusion of at least the antigen binding region of an antibody of the present disclosure and a serum protein (e.g., human serum albumin or a fragment thereof) to increase the half-life of the resulting molecule. Such a process is described, for example, in EP 0322094, the contents of which are hereby incorporated by reference.

Another possibility is the fusion of at least the antigen binding region of an antibody of the present disclosure with a protein capable of binding to a serum protein (such as human serum albumin) to prolong the half-life of the resulting molecule. Such a process is described, for example, in EP 0486525, the contents of which are hereby incorporated by reference.

Nucleic acid molecules encoding antibodies of the disclosure

Another aspect of the disclosure relates to nucleic acid molecules encoding the antibodies of the disclosure. The term "nucleic acid" is used interchangeably herein with the term "polynucleotide" and refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single-or double-stranded form. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, have similar binding properties as the reference nucleic acid, and are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, but are not limited to, phosphorothioate, phosphoramidate, methylphosphonate, chiral-methylphosphonate, 2-O-methyl ribonucleotide, peptide-nucleic acid (PNA). In some embodiments, the nucleic acid can be mRNA.

Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences as well as the sequence explicitly indicated. In particular, degenerate codon substitutions may be obtained by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed base and/or deoxyinosine residues (see: Batzer et al, Nucleic Acids Res [ Nucleic Acids research ] 1991; 25(19): 5081; Ohtsuka et al, J Biol Chem [ J. Biol. Chem ] 1985; 260(5): 2605-8; Rossolini et al, Mol Cell Probes [ molecular and cellular Probes ] 1994; 8(2): 91-8; the contents of each of which are hereby incorporated by reference.

Exemplary full-length heavy and light chain nucleotide sequences of anti-NPR 1 antibodies are provided herein. In some embodiments, the nucleic acid molecule is one or more of the nucleic acid molecules identified in table 2, e.g., a nucleic acid molecule encoding an anti-NPR 1 antibody or antigen-binding fragment thereof. In some other embodiments, the nucleic acid molecules described herein comprise nucleotide sequences that are substantially identical (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) to the nucleotide sequences of those nucleic acid molecules identified in table 2. The polypeptides encoded by these polynucleotides are capable of binding to NPR1 protein (e.g., human NPR1) when expressed from an appropriate expression vector.

Also provided herein are polynucleotides encoding at least one CDR region and typically all three CDR regions of the heavy and/or light chain from an anti-NPR 1 antibody or antigen-binding fragment of the disclosure. Further provided herein are polynucleotides encoding all or substantially all of the variable region sequences of the heavy and/or light chains of an exemplary anti-NPR 1 antibody or antigen-binding fragment of the disclosure. Due to the degeneracy of the genetic code, a variety of nucleic acid sequences will encode each of the immunoglobulin amino acid sequences.

In some embodiments, the nucleic acid molecules disclosed herein encode the variable and constant regions of an antibody. In some embodiments, a nucleic acid molecule disclosed herein comprises nucleotides encoding a full length heavy chain sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) to a heavy chain sequence of one of the antibodies described herein, including those sequences in table 2. In some embodiments, a nucleic acid molecule disclosed herein comprises nucleotides encoding a full length light chain sequence that is substantially identical (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) to a light chain sequence of one of the antibodies described herein, including those sequences in table 2.

These nucleic acids may be present in intact cells, in cell lysates, or may be nucleic acids in partially purified or substantially pure form. Nucleic acids are "isolated" or "become substantially pure" when purified from other cellular components or other contaminants (e.g., other cellular nucleic acids or proteins) by standard techniques, including alkali/SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and other techniques well known in the art. See, e.g., Ausubel et al (eds.), 1987Current Protocols in Molecular Biology [ Molecular Biology laboratory Manual ], Greene Publishing and Wiley Interscience [ Green publication and Weley Cross sciences ], New York, the contents of which are hereby incorporated by reference. The nucleic acids of the disclosure may be, for example, DNA or RNA, and may or may not comprise intron sequences. In one embodiment, the nucleic acid is a cDNA molecule. The nucleic acid may be present in a vector, such as a phage display vector, or in a recombinant plasmid vector.

Nucleic acids of the disclosure can be obtained using standard molecular biology techniques. For antibodies expressed by a hybridoma (e.g., a hybridoma prepared from a transgenic mouse carrying human immunoglobulin genes, as described further herein), cdnas encoding the light and heavy chains of the antibody produced by the hybridoma can be obtained by standard PCR amplification or cDNA cloning techniques. For antibodies obtained from immunoglobulin gene libraries (e.g., using phage display technology), antibody-encoding nucleic acids can be recovered from a variety of phage clones that are members of the library.

Polynucleotide sequences can be generated by de novo solid phase DNA synthesis or by PCR mutagenesis of existing sequences (e.g., as described herein, e.g., in table 2). Direct chemical synthesis of nucleic acids can be accomplished by methods known in the art, for example, Narang et al, 1979, meth.enzymol. [ methods of enzymology ]68: 90; the phosphodiester method of Brown et al, meth.enzymol. [ methods of enzymology ]68:109,1979; the diethylphosphoramidite method of Beaucage et al, tetra.Lett. [ tetrahedron letters ],22:1859,1981; and U.S. Pat. No. 4,458,066 (the respective contents of which are hereby incorporated by reference). The introduction of mutations into polynucleotide sequences by PCR can be carried out as described in, for example, PCR Technology: Principles and Applications for DNA Amplification [ PCR Technology: principles and applications for DNA amplification ], h.a. erlich (editors), frieman Press, new york (Freeman Press, NY, n.y.), 1992; PCR Protocols A Guide to Methods and Applications [ PCR protocol: methods and application guidelines ], Innis et al, (eds.), Academic Press, San Diego, Calif., 1990; mattila et al, Nucleic Acids Res. [ Nucleic acid research ]19:967,1991; and Eckert et al, PCR Methods and Applications [ PCR Methods and Applications ]1:17, 1991.

Once the DNA fragments encoding the VH and VL segments are obtained, these DNA fragments can be further manipulated by standard recombinant DNA techniques, for example to convert the variable region genes to full-length antibody chain genes, to Fab fragment genes, or to scFv genes. In these manipulations, a DNA fragment encoding a VL or VH is operably linked to another DNA molecule, or to a fragment encoding another protein (e.g., an antibody constant region or flexible linker). As used in this context, the term "operably linked" is intended to mean that two DNA fragments are joined in a functional manner, e.g., such that the amino acid sequences encoded by the two DNA fragments are held in frame, or such that the protein is expressed under the control of a desired promoter.

Isolated DNA encoding the VH region can be converted to a full-length heavy chain gene by operably linking the DNA encoding the VH to another DNA molecule encoding the heavy chain constant region (CH1, CH2, and CH 3). The sequence of the human heavy chain constant region gene is known in the art (see, e.g., Kabat, e.a. et al, 1991Sequences of Proteins of Immunological Interest [ immune-related protein Sequences ], fifth edition, department of health and welfare, NIH publication No. 91-3242, the contents of which are hereby incorporated by reference), and DNA fragments encompassing these regions can be obtained by standard PCR amplification. The heavy chain constant region may be an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, or IgD constant region. In some embodiments, the heavy chain constant region is an IgG1 isotype. For Fab fragment heavy chain genes, the DNA encoding VH can be operably linked to another DNA molecule encoding only the heavy chain CH1 constant region.

The isolated DNA encoding the VL region can be converted to a full-length light chain gene (and to a Fab light chain gene) by operably linking the DNA encoding the VL to another DNA molecule encoding the light chain constant region CL. The sequence of the human light chain constant region gene is known in the art (see, e.g., Kabat, e.a. et al, 1991Sequences of Proteins of Immunological Interest [ immune-related protein Sequences ], fifth edition, department of health and welfare, NIH publication No. 91-3242, the contents of which are hereby incorporated by reference), and DNA fragments encompassing these regions can be obtained by standard PCR amplification. The light chain constant region can be a kappa or lambda constant region.

To generate the scFv genes, the VH and VL encoding DNA fragments are operably linked to another fragment encoding a flexible linker such that the VH and VL sequences can be expressed as a continuous single-stranded protein whose VL and VH regions are linked by a flexible linker (see, e.g., Bird et al, 1988, Science 242:423 + 426; Huston et al, 1988Proc. Natl. Acad. Sci. USA 85:5879 + 5883; McCafferty et al, 1990, Nature [ Nature ]348:552 + 554; the contents of each of which are hereby incorporated by reference).

Carrier

A variety of expression vectors can be used to express a polynucleotide encoding an antibody or antigen-binding fragment thereof of the disclosure. Both viral-based and non-viral expression vectors can be used to produce antibodies in mammalian host cells. Non-viral vectors and systems include plasmids, episomal vectors (typically having expression cassettes for expression of proteins or RNA), and human artificial chromosomes (see, e.g., Harrington et al, Nat Genet. [ natural genetics ]15:345,1997, the contents of which are hereby incorporated by reference). For example, non-viral vectors useful for expressing polynucleotides and polypeptides of multispecific antibodies or domains thereof of the present disclosure in mammalian (e.g., human) cells include pThioHis a, pThioHis B, and pThioHis C, pcdna3.1/His, pEBVHis a, pEBVHis B, and pEBVHis C (Invitrogen, San Diego, Calif.), MPS V vector, and a variety of other vectors known in the art for expressing other proteins. Useful viral vectors include retroviral, adenoviral, adeno-associated viral, herpes virus based vectors, SV40, papilloma virus, HBP EB virus, vaccinia virus vectors and Semliki Forest Virus (SFV) based vectors. See, Brent et al, supra; smith, annu.rev.microbiol. [ microbiological annual review ]49:807,1995; and Rosenfeld et al, Cell [ Cell ]68:143,1992, the contents of each of which are hereby incorporated by reference.

The choice of expression vector will depend on the intended host cell in which the vector is to be expressed. Expression vectors for mammalian host cells can include expression control sequences such as origins of replication, promoters and enhancers (see, e.g., Queen et al, immunol. rev. [ immunologic review ]89:49-68,1986, the contents of which are hereby incorporated by reference) and necessary processing information sites such as ribosome binding sites, RNA splice sites, polyadenylation sites, and transcription terminator sequences. These expression vectors typically contain promoters derived from mammalian genes or from mammalian viruses. Suitable promoters may be constitutive, cell type specific, stage specific and/or regulatable. Useful promoters include, but are not limited to, the metallothionein promoter, the constitutive adenovirus major late promoter, the dexamethasone-inducible MMTV promoter, the SV40 promoter, the MRP poiiiii promoter, the constitutive MPS V promoter, the tetracycline-inducible CMV promoter (e.g., the human i.e., early CMV promoter), the constitutive CMV promoter, and known promoter-enhancer combinations.

The culture of the transformed organism can be expanded under non-inducing conditions without biasing the population of host cells to better tolerate the coding sequences of their expression products. In addition to a promoter, other regulatory elements may be required or desired for efficient expression of the antibodies or fragments thereof of the present disclosure. These elements typically include the ATG initiation codon and adjacent ribosome binding sites or other sequences. Furthermore, expression efficiency can be increased by including enhancers suitable for the cell system in use (see, e.g., Scharf et al, Results Probl. cell Differ. [ Results and problems in cell differentiation ]20:125,1994; and Bittner et al, meth. enzymol. [ methods of enzymology ],153:516, 1987; the contents of each of which are hereby incorporated by reference). For example, the SV40 enhancer or the CMV enhancer may be used to increase expression in a mammalian host cell.

Accordingly, the present disclosure provides cloning or expression vectors comprising one or more of the nucleic acid sequences of the antibodies set forth in table 2. Further, the disclosure provides a cloning or expression vector comprising a nucleic acid encoding one or more of the nucleotide sequences set forth in table 2.

Host cell

For expression of the light and heavy chains, one or more expression vectors encoding the heavy and light chains can be transferred into a host cell by standard techniques.

The method used to introduce the expression vector containing the polynucleotide sequence of interest varies depending on the type of cellular host. For example, calcium chloride transfection is commonly used for prokaryotic cells, while calcium phosphate treatment or electroporation may be used for other cellular hosts. (see generally Sambrook et al, supra, the contents of which are hereby incorporated by reference). Other methods include, for example, electroporation, calcium phosphate treatment, liposome-mediated transformation, injection and microinjection, impact, virosomes, immunoliposomes, polycationic nucleic acid conjugates, naked DNA, artificial virions, fusions with the herpes virus structural protein VP22 (Elliot and O' Hare, Cell [ Cell ]88:223,1997, the contents of which are hereby incorporated by reference), agent-enhanced DNA uptake and ex vivo transduction.

Expression of the antibodies of the disclosure in prokaryotic or eukaryotic host cells is theoretically possible. Expression of antibodies in eukaryotic cells (particularly in mammalian host cells) is discussed, as such eukaryotic cells (and particularly mammalian cells) are more likely to assemble and secrete properly folded and immunologically active antibodies than prokaryotic cells. Prokaryotic expression of antibody genes has been reported to be ineffective for producing active antibodies in high yields (Boss, M.A. and Wood, C.R.,1985Immunology Today 6:12-13, the contents of which are hereby incorporated by reference).

For long-term high-yield production of recombinant proteins, stable expression is often desired. For example, cell lines stably expressing an antibody or antigen-binding fragment thereof of the present disclosure can be prepared using expression vectors of the present disclosure containing viral origins of replication or endogenous expression elements and a selectable marker gene. After introducing the vector, the cells can be grown in enriched medium for 1-2 days and then switched to selective medium. The purpose of the selectable marker is to confer resistance to selection and its presence allows the growth of cells that successfully express the introduced sequence in a selective medium. Resistant, stably transfected cells can be propagated using tissue culture techniques appropriate to the cell type. Accordingly, the disclosure provides methods of producing an antibody or antigen-binding fragment of the disclosure, wherein the methods comprise the step of culturing a host cell comprising a nucleic acid encoding the antibody or antigen-binding fragment.

In some embodiments, the anti-NPR 1 antibodies or antigen-binding fragments of the disclosure are expressed and produced using mammalian host cells. For example, they may be hybridoma cell lines expressing endogenous immunoglobulin genes or mammalian cell lines containing exogenous expression vectors. These include any normally non-immortalized or normal or abnormal immortalized animal or human cell. For example, a number of suitable host cell lines capable of secreting intact immunoglobulins have been developed, including CHO cell lines, various COS cell lines, HeLa cells, myeloma cell lines, transformed B cells, and hybridomas. Exemplary host cells include, but are not limited to, Chinese Hamster Ovary (CHO) cells, Human Embryonic Kidney (HEK) cells (e.g., HEK293T, HEK293F), monkey kidney (COS) cells (e.g., COS-1, COS-7), Baby Hamster Kidney (BHK) cells (e.g., BHK-21), African green monkey kidney cells (e.g., BSC-1), HeLa cells, human hepatocellular carcinoma cells (e.g., Hep G2), myeloma cells (e.g., NS0, 653, SP2/0), lymphoma cells, oocytes, and cells from transgenic animals (e.g., mammary epithelial cells), or any derived, immortalized or transformed cells thereof. In particular, for use with NS0 myeloma cells, another expression system is the GS gene expression system shown in WO 87/04462, WO 89/01036 and EP 0338841, the respective contents of which are hereby incorporated by reference. Upon introduction of a recombinant expression vector encoding an antibody nucleic acid into a mammalian host cell, the antibody is produced by culturing the host cell for a period of time sufficient to allow expression of the antibody in the host cell or secretion of the antibody into the medium in which the host cell is grown. The antibody can be recovered from the culture medium using standard protein purification methods. Such purified antibodies of the present disclosure may be used for any purpose, including but not limited to the methods and uses described herein, and/or as part of a pharmaceutical composition as described herein.

In a further alternative, the host cell may be a yeast or filamentous fungus engineered for a mammalian-like glycosylation pattern and capable of producing antibodies lacking fucose as a glycosylation pattern (see, e.g., EP 1297172, the contents of which are hereby incorporated by reference).

Accordingly, the disclosure provides a host cell comprising one or more of the vectors or nucleic acid sequences of the disclosure described above.

Generation of monoclonal antibodies of the disclosure

Monoclonal antibodies (mAbs) can be produced by a variety of techniques including conventional monoclonal antibody methods, such as the somatic hybridization technique of Kohler and Milstein,1975Nature [ Nature ]256:495 (the contents of which are hereby incorporated by reference). Many techniques for generating monoclonal antibodies can be used, for example, viral or oncogenic transformation of B lymphocytes.

Hybridomas can be prepared using, for example, the murine system. The immunization protocol and isolation of immune splenocytes for fusion can be performed according to any suitable procedure. Chimeric or humanized antibodies can be prepared based on the sequence of a murine monoclonal antibody prepared as described herein. DNA encoding the heavy and light chain immunoglobulins can be obtained from a murine hybridoma of interest and engineered to contain non-murine (e.g., human) immunoglobulin sequences using standard molecular biology techniques. For example, to produce chimeric antibodies, the murine variable regions can be linked to human constant regions using any known method (see, e.g., U.S. Pat. No. 4,816,567, the contents of which are hereby incorporated by reference). To generate humanized antibodies, the murine CDR regions can be inserted into a human framework using any known method. See, for example, US 5225539 and US patent numbers 5530101, 5585089, 5693762 and 6180370, the respective contents of which are hereby incorporated by reference.

Transgenic or transchromosomal mice carrying portions of the human immune system rather than the mouse system can also be used to produce human monoclonal antibodies. These transgenic and transchromosomal mice are included hereinRespectively called HuMAb

And KM mice, and collectively referred to herein as "human Ig mice".

HuMAb

(Meddarrax, Inc.) contains a human immunoglobulin gene minilocus (minioci) encoding unrearranged human heavy (mu and gamma) and K light chain immunoglobulin sequences, as well as targeted mutations that inactivate endogenous mu and K chain loci (see, e.g., Lonberg et al, 1994Nature [ Nature ] (Medarex, Inc.)]368 (368), (6474) 856-. Thus, mice show reduced expression of mouse IgM or K, and in response to immunization, the introduced human heavy and light chain transgenes undergo class switching and somatic mutation to generate high affinity human IgG_KMonoclonal (Lonberg, N.et al, 1994 supra; in Lonberg, N.1994 Handbook of Experimental Pharmacology]113: 49-101; lonberg, n. and huskzar, d.,1995 inter.rev.immunol. [ international reviews of immunology [ ]]65-93 and Harding, F. and Lonberg, N.,1995Ann.N.Y.Acad.Sci. [ New York scientific academic annual newspaper ] ]764:536-546, the contents of each of which are hereby incorporated by reference). HuMAb

The preparation and use of and the genomic modifications made by such mice are further described in the following documents: taylor, L. et al, 1992Nucleic Acids Research [ Nucleic Acids Research]6287-6295; chen, J. et al, 1993International Immunology [ International Immunology]647-656; tuaillon et al, 1993Proc.Natl.Acad.Sci.USA [ Proc. Natl. Acad. Sci.]3720 and 3724; choi et al, 1993Nature Genetics [ Nature Genetics]4: 117-; chen, J. et al, 1993EMBO J. [ J. European society for molecular biology]12: 821-; tuaillon et al, 1994J.Immunol. [ J.Immunol. ]]152: 2912-2920; taylor, L. et al, 1994International Immunology]579-; and Fishwild, D. et al, 1996Nature BIo technology (Natural Biotechnology)]14:845-851, the respective contents of which are hereby incorporated by reference. See also, U.S. Pat. nos. 5,545,806, 5,569,825, 5,625,126, 5,633,425, 5,789,650, 5,877,397, 5,661,016, 5,814,318, 5,874,299, and 5,770,429, US 5,545,807, WO 92/103918, WO 93/12227, WO 94/25585, WO 97/113852, WO 98/24884 and WO 99/45962, and WO 01/14424; the contents of each are hereby incorporated by reference.

In another example, human antibodies can be produced using mice carrying human immunoglobulin sequences on transgenes and transchromosomes, such as mice carrying human heavy chain transgenes and human light chain transchromosomes. Such mice, referred to herein as "KM mice", are described in detail in WO 02/43478, the contents of which are hereby incorporated by reference.

Still further, alternative transgenic animal systems expressing human immunoglobulin genes are available in the art and can be used to produce antibodies of the present disclosure. For example, an alternative transgene system known as Xenomouse (Abgenix, Inc.) may be used. Such mice are described, for example, in U.S. Pat. nos. 5,939,598, 6,075,181, 6,114,598, 6,150,584, and 6,162,963, the contents of each of which are hereby incorporated by reference.

In addition, alternative transgenic animal systems expressing human immunoglobulin genes are available in the art and can be used to produce antibodies of the present disclosure. For example, a mouse known as a "TC mouse" carrying a human heavy chain transchromosome and a human light chain transchromosome may be used; such mice are described in Tomizuka et al, 2000Proc. Natl. Acad. Sci. USA [ Proc. Natl. Acad. Sci ]97:722-727, the contents of which are hereby incorporated by reference. Furthermore, cattle carrying human heavy and light chain transchromosomes have been described in the art (Kuroiwa et al, 2002Nature Biotechnology [ Nature Biotechnology ]20:889-894, the contents of which are hereby incorporated by reference) and can be used to produce antibodies of the present disclosure.

Human monoclonal antibodies of the disclosure can also be prepared using phage display methods directed to screening human immunoglobulin gene libraries. Such phage display methods for isolating human antibodies have been established in the art or are described in the examples below. See, for example: U.S. Pat. nos. 5,223,409, 5,403,484, and 5,571,698, U.S. Pat. nos. 5,427,908 and 5,580,717, U.S. Pat. nos. 5,969,108 and 6,172,197, and U.S. Pat. nos. 5,885,793, 6,521,404, 6,544,731, 6,555,313, 6,582,915, and 6,593,081, the respective contents of which are hereby incorporated by reference.

Human antibodies or antigen-binding fragments of the disclosure can also be prepared using SCID mice into which human immune cells have been reconstituted so that upon immunization a human antibody response can be generated. Such mice are described, for example, in U.S. patent nos. 5,476,996 and 5,698,767, the contents of each of which are hereby incorporated by reference.

Antibodies of the disclosure can be prepared by any of the methods described herein.

Generation of hybridomas that produce antibodies or antigen-binding fragments of the disclosure

To generate hybridomas that produce antibodies or antigen-binding fragments of the present disclosure, spleen cells and/or lymph node cells from immunized mice can be isolated and fused with an appropriate immortalized cell line (e.g., a mouse myeloma cell line). The resulting hybridomas can be screened for the production of antigen-specific antibodies. For example, a single cell suspension of splenic lymphocytes from immunized mice can be fused with one-sixth the number of P3X63-Ag8.653 non-secreting mouse myeloma cells (ATCC, CRL 1580) with 50% PEG. Cells were plated at approximately 2X 145 onto flat-bottomed microtiter plates, followed by two weeks of incubation in selective medium containing 20% fetal clone serum, 18% "653" conditioned medium, 5% trioxazocine (origen) (IGEN), 4mM L-glutamine, 1mM sodium pyruvate, 5mM HEPES, 0.055mM 2-mercaptoethanol, 50 units/ml penicillin, 50mg/ml streptomycin, 50mg/ml gentamicin, and 1 XHAT (Sigma; HAT was added at 24 hours post-fusion). After about two weeks, the cells can be cultured in medium in which HAT is replaced with HT. Individual wells can then be screened by ELISA for human monoclonal IgM and IgG antibodies. Once extensive hybridoma growth has occurred, the medium can generally be observed after 10-14 days. Antibody-secreting hybridomas can be replated, screened again, and if still positive for human IgG, these monoclonal antibodies can be subcloned at least twice by limiting dilution. The stable subclones can then be cultured in vitro to produce small amounts of antibody in tissue culture medium for characterization.

To purify the antibody or antigen-binding fragment thereof, selected hybridomas are grown in two-liter spinner flasks for antibody purification. The supernatant may be filtered and concentrated before affinity chromatography on protein a-sepharose (Pharmacia, Piscataway, n.j.) is performed. The eluted IgG can be checked by gel electrophoresis and high performance liquid chromatography to ensure purity. The buffer solution can be replaced by PBS and can pass through OD₂₈₀The concentration was determined using an extinction coefficient of 1.43. The antibody or antigen binding fragment can be aliquoted and stored at-80 ℃.

Hybridomas that produce antibodies or antigen-binding fragments of the disclosure can be produced, for example, using the methods described herein.

Generation of transfectomas that produce antibodies or antigen-binding fragments of the disclosure

Antibodies or antigen-binding fragments of the disclosure can also be produced in host cell transfectomas using, for example, a combination of suitable recombinant DNA techniques and gene transfection methods (e.g., Morrison, S. (1985) Science 229:1202, the contents of each of which are hereby incorporated by reference).

For example, to express an antibody or antigen-binding fragment thereof, DNA encoding partial or full-length light and heavy chains can be obtained by standard molecular biology techniques (e.g., PCR amplification or cDNA cloning using a hybridoma expressing the antibody of interest), and these DNAs can be inserted into expression vectors such that the genes are operably linked to transcriptional and translational control sequences. In this context, the term "operably linked" is intended to mean that the antibody gene is linked into a vector such that transcriptional and translational control sequences within the vector exert their effect of regulating transcription of the antibody gene And the intended function of the translation. The expression vector and expression control sequences are selected to be compatible with the expression host cell used. The antibody light chain gene and the antibody heavy chain gene may be inserted into separate vectors, or both genes may be inserted into the same expression vector. The antibody gene is inserted into the expression vector by standard methods (e.g., linking the antibody gene fragment to complementary restriction sites on the vector, or blunt-ended if no restriction sites are present). The light and heavy chain variable regions of the antibodies described herein can be used to generate full-length antibody genes of any antibody isotype by: these light and heavy chain variable regions are inserted into expression vectors that already encode the heavy and light chain constant regions of the desired isotype, such that V_HSegments are operably linked to one or more CH segments within a vector, and V_LThe segments are operably connected to CL segments within the carrier. Additionally or alternatively, the recombinant expression vector may encode a signal peptide that facilitates secretion of the antibody chain from the host cell. The antibody chain gene can be cloned into a vector such that the signal peptide is linked in-frame to the amino terminus of the antibody chain gene. The signal peptide may be an immunoglobulin signal peptide or a heterologous signal peptide (i.e., a signal peptide from a non-immunoglobulin protein).

In addition to the antibody chain gene, the recombinant expression vectors of the disclosure carry regulatory sequences that control the expression of the antibody chain gene in the host cell. The term "regulatory sequence" is intended to include promoters, enhancers and other expression control elements (e.g., polyadenylation signals) that control the transcription or translation of antibody chain genes. Such regulatory sequences are described, for example, in Goeddel (Gene Expression Technology [ Gene Expression Technology ]; Methods in Enzymology [ Methods of Enzymology ]185, Academic Press [ Academic Press ], 1990, san Diego, Calif., each of which is hereby incorporated by reference). It will be appreciated by those skilled in the art that the design of the expression vector (including the choice of regulatory sequences) may depend on factors such as: selection of the host cell to be transformed, expression level of the desired protein, etc. Regulatory sequences for expression in mammalian host cells include viral elements that direct expression of high levels of proteins in mammalian cells, such as promoters and/or enhancers derived from Cytomegalovirus (CMV), simian virus 40(SV40), adenoviruses (e.g., adenovirus major late promoter (AdMLP)), and polyoma. Alternatively, non-viral regulatory sequences may be used, such as the ubiquitin promoter or the P-globulin promoter. Still further, the regulatory elements are composed of sequences from different sources, such as the SRa promoter system, which contains sequences from the SV40 early promoter as well as the long terminal repeat of the human T cell leukemia virus type 1 (Takebe, Y. et al, 1988mol. cell. biol. [ molecular cell biology ]8: 466-.

In addition to antibody chain genes and regulatory sequences, recombinant expression vectors of the disclosure may carry additional sequences, such as sequences that regulate replication of the vector in a host cell (e.g., an origin of replication) and a selectable marker gene. Selectable marker genes facilitate the selection of host cells into which a vector has been introduced (see, e.g., U.S. Pat. nos. 4,399,216, 4,634,665, and 5,179,017, the contents of each of which are hereby incorporated by reference). For example, selectable marker genes typically confer resistance to drugs such as G418, hygromycin or methotrexate to host cells into which the vector has been introduced. Selectable marker genes include the dihydrofolate reductase (DHFR) gene (used in DHFR-host cells for methotrexate selection/amplification) and the neo gene (used for G418 selection).

For expression of the light and heavy chains, one or more expression vectors encoding the heavy and light chains are transfected into the host cell by standard techniques. The term "transfection" in different forms is intended to encompass a variety of techniques commonly used for introducing foreign DNA into prokaryotic or eukaryotic host cells, such as electroporation, calcium phosphate precipitation, DEAE-dextran transfection, and the like. Expression of the antibodies of the disclosure in prokaryotic or eukaryotic host cells is theoretically possible. Expression of antibodies in eukaryotic cells (particularly in mammalian host cells) is discussed, as such eukaryotic cells (and particularly mammalian cells) are more likely to assemble and secrete properly folded and immunologically active antibodies than prokaryotic cells. Prokaryotic expression of antibody genes has been reported to be ineffective for producing active antibodies in high yields (Boss, M.A. and Wood, C.R.,1985Immunology Today 6:12-13, the contents of which are hereby incorporated by reference).

Described elsewhere herein are mammalian host cells for expressing the antibodies of the disclosure. Upon introduction of a recombinant expression vector encoding a gene for an antibody into a mammalian host cell, the antibody is produced by culturing the host cell for a period of time sufficient to allow expression of the antibody in the host cell or secretion of the antibody into the medium in which the host cell is grown. The antibody can be recovered from the culture medium using standard protein purification methods. Accordingly, the disclosure provides methods for producing an anti-NPR 1 antibody or antigen-binding fragment thereof of the disclosure, the methods comprising culturing a host cell of the disclosure and isolating the antibody or antigen-binding fragment thereof.

Therapeutic uses and methods

Method of treatment

Provided herein are methods of treating diseases associated with loss of NPR1 function by using an anti-NPR 1 antibody or antigen-binding fragment thereof disclosed herein (e.g., an antibody or group of antibodies as defined in table 2, table 3, or table 4). In some embodiments, the antibody or antigen-binding fragment thereof can be selected from WW _ LALA, XX _ DAPA, XX _ N30 _ DAPA, XX _ LALA, XX _ dala, XX _ DAPA, XX _ LALA, XX _ DAPA, XX _ N30 _ DAPA, XX _ LALA, XX _ dala, zzlala, XX _ dala, YY _ LALA, and zzla.

In some embodiments, the antibody or antigen binding fragment thereof may be selected from WW01_ LALA, WW03_ LALA, XX01_ LALA, XX01_ DAPA, XX01_ N30S _ DAPA, XX03_ LALA, XX04_ LALA, XX06_ LALA, XX06_ DAPA, XX07_ LALA, XX08_ LALA, XX08_ DAPA, XX08_ N30S _ DAPA, XX08_ N30Q _ DAPA, XX09_ LALA, XX11_ LALA, XX12_ LALA, XX13_ LALA, XX14_ DAPA, XX14_ laja, 14_ lazzla, 14_ dala, 14_ laxx _ lay, 14_ layla, 14_ laxx _ lay3672 _ lay, 14_ layy, 14_ layla, 14_ laxx _ layy, 14_ layla, 14_ lay3672 _ lay, 14_ laxx _ layy, 14_ laylay, 14_ lay, 14_ lay3672, 14_ laylay, 14_ lay, 14_ laylay _ lay _ laxx _ lay, 14_ lay _ laxx 14, 14_ lay _ laxx _ lay _ laxx, 14, 36yy _ lay. In some embodiments, the antibody or antigen-binding fragment thereof may be selected from WW05_ LALA, WW06_ LALA, YY05_ LALA, YY06_ LALA, YY07_ LALA, ZZ05_ LALA, ZZ14_ LALA, and ZZ16_ LALA. In some embodiments, the antibody or antigen-binding fragment thereof may be XX16_ DAPA. In some embodiments, the antibody or antigen-binding fragment thereof may be XX16_ LALA.

In some embodiments, the disease associated with loss of NPR1 function is a cardiovascular disorder. In some embodiments, the cardiovascular disorder is selected from: hypertension, peripheral vascular disease, heart failure, Coronary Artery Disease (CAD), Ischemic Heart Disease (IHD), mitral stenosis and regurgitation, angina, hypertrophic cardiomyopathy, diabetic cardiomyopathy, supraventricular and ventricular arrhythmias, arrhythmia, Atrial Fibrillation (AF), new atrial fibrillation, recurrent atrial fibrillation, cardiac fibrosis, atrial flutter, detrimental vascular remodeling, plaque stabilization and Myocardial Infarction (MI). In some embodiments, the disease associated with loss of NPR1 function is heart failure, Hypertrophic Cardiomyopathy (HCM), hypertension, preeclampsia, asthma, glaucoma, or cytokine release syndrome. In some embodiments, the heart failure is selected from the group consisting of heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), heart failure after acute myocardial infarction, or acute compensated heart failure. In some embodiments, the hypertrophic cardiomyopathy is ventricular hypertrophy. In some embodiments, the hypertension is selected from refractory hypertension, hypertensive heart disease, pulmonary hypertension, pulmonary arterial hypertension, isolated systolic hypertension, refractory hypertension, and pulmonary arterial hypertension. In some embodiments, the hypertension is selected from refractory hypertension and hypertensive heart disease.

In some embodiments, the disease associated with loss of NPR1 function is a renal disorder. In some embodiments, the renal disorder is selected from: diabetic renal insufficiency, non-diabetic renal insufficiency, renal failure, diabetic nephropathy, non-diabetic nephropathy, acute kidney injury, contrast-induced nephropathy, nephrotic syndrome, glomerulonephritis, scleroderma, glomerulosclerosis, primary nephrotic proteinuria, renovascular hypertension, diabetic retinopathy and end-stage renal disease (ESRD), endothelial dysfunction, diastolic dysfunction, renal fibrosis and Polycystic Kidney Disease (PKD).

NPR 1-related disorders also include any other disorder directly or indirectly associated with aberrant NPR1 activity and/or expression. Also provided herein are methods of treating NPR 1-associated disorders directly or indirectly associated with aberrant NPR1 activity and/or expression by using an anti-NPR 1 antibody or antigen-binding fragment disclosed herein (e.g., from table 2, table 3, or table 4, e.g., XX16_ DAPA or XX16_ LALA).

In some embodiments, the disclosure provides a method of treating an undesirable condition, disease, or disorder associated with natriuretic peptide receptor activity in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of an antibody or antigen binding fragment disclosed herein. In some embodiments, the disclosure provides for the use of an antibody or antigen-binding fragment disclosed herein to treat an undesirable condition, disease, or disorder associated with natriuretic peptide receptor activity in a subject in need thereof. In some embodiments, the disclosure provides an antibody or antigen-binding fragment disclosed herein for use in a method of treating an undesirable condition, disease, or disorder associated with natriuretic peptide receptor activity. In some embodiments, the disclosure provides an antibody or antigen-binding fragment disclosed herein for use in the manufacture of a medicament for treating an undesirable condition, disease, or disorder associated with natriuretic peptide receptor activity. Such conditions, diseases and disorders include, but are not limited to, cardiovascular disorders (e.g., hypertension, peripheral vascular disease, heart failure (including, but not limited to, heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), heart failure after acute myocardial infarction, or acute compensatory heart failure), coronary heart disease (CAD), Ischemic Heart Disease (IHD), mitral stenosis and regurgitation, angina, hypertrophic cardiomyopathy (e.g., ventricular hypertrophy), diabetic cardiomyopathy, supraventricular and ventricular arrhythmias, arrhythmia, Atrial Fibrillation (AF), new atrial fibrillation, recurrent atrial fibrillation, cardiac fibrosis, atrial flutter, harmful vascular remodeling, plaque stabilization, or Myocardial Infarction (MI)), hypertension (refractory hypertension, hypertensive heart disease, pulmonary hypertension, isolated systolic hypertension, Myocardial Infarction (MI)), hypertension (refractory hypertension, peripheral vascular disease, heart failure including, but not limited to, stroke-induced myocardial infarction (stroke-induced stroke), stroke-induced stroke, stroke-induced stroke-induced stroke-induced stroke-induced stroke-induced stroke induced, Refractory hypertension or pulmonary hypertension), preeclampsia, asthma, glaucoma, cytokine release syndrome, and/or renal disorders (e.g., diabetic renal insufficiency, non-diabetic renal insufficiency, renal failure, diabetic nephropathy, non-diabetic nephropathy, acute kidney injury, contrast-induced nephropathy, nephrotic syndrome, glomerulonephritis, scleroderma, glomerulosclerosis, primary nephrotic proteinuria, renovascular hypertension, diabetic retinopathy and end-stage renal disease (ESRD), endothelial dysfunction, diastolic dysfunction, renal fibrosis and Polycystic Kidney Disease (PKD)).

In some embodiments, such methods comprise administering to a subject in need of treatment a therapeutically effective amount of an antibody, or antigen-binding fragment thereof, that specifically binds to the same epitope as one of the antibodies described herein. For example, such methods comprise administering to a subject in need of treatment a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to the same epitope as XX 16. In another embodiment, such methods comprise administering to a subject in need of treatment a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to the same epitope as WW 03. In another embodiment, such methods comprise administering to a subject in need of treatment a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds to the same epitope as WW 06.

All the aforementioned embodiments of the protection and treatment method according to the invention are equally applicable

Use of any of the antibodies or antigen binding fragments as described herein in the manufacture of a medicament for use according to the invention,

According to the invention of the antibody or antigen binding fragment in any one of the use,

any of the antibodies or antigen-binding fragments described herein for use according to the invention,

a pharmaceutical composition comprising any of the antibodies or antigen-binding fragments described herein for use according to the invention,

use of a pharmaceutical composition comprising any of the antibodies or antigen-binding fragments described herein according to the invention, and

use of a pharmaceutical composition comprising any of the antibodies or antigen-binding fragments described herein in the manufacture of a medicament for use according to the invention.

Combination therapy

The various treatments described above may be combined with other therapeutic partners (partners) or therapeutic agents, such as the current standard of care for diseases associated with loss of function of NPR1, e.g., the current standard of care for one or more of the diseases or disorders discussed herein. For example, the NPR1 antibodies or antigen-binding fragments thereof described herein can be combined with one or more of an ACE (angiotensin converting enzyme) inhibitor, an Angiotensin Receptor Blocker (ARB), an enkephalinase inhibitor, a beta blocker, a diuretic, a calcium channel blocker, a cardiac glycoside, a sodium-glucose co-transporter 2 inhibitor (SGLT2i), or a combination thereof. As non-limiting examples, the NPR1 antibody or antigen-binding form may be combined with an additional therapeutic agent selected from the group consisting of: enalapril, benazepril, captopril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, trandolapril, valsartan, azilsartan, candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, sacubitril, bisoprolol, carvedilol, propranolol, metoprolol tartrate, metoprolol succinate, thiazide diuretics, cyclic diuretics, potassium sparing diuretics, amlodipine, clevidipine, diltiazem, felodipine, isradipine, nicardipine, nifedipine, nisoldipine, lisinopril, Verapamil, digitoxin, canagliflozin, dapagliflozin, eprazigliflozin, eggliflozin, and combinations thereof. Exemplary diuretics and digitoxins include, but are not limited to, chlorothiazide, chlorthalidone, hydrochlorothiazide, indapamide, metolazone, bumetanide, ethacrynic acid, furosemide, torasemide, amiloride, eplerenone, spironolactone, triamterene, digoxin, and combinations thereof. In some embodiments, the NPR1 antibodies or antigen-binding fragments thereof described herein can be combined with an angiotensin receptor-enkephalinase inhibitor (ARNi), such as a combination of sabotary and valsartan (e.g., as described herein)

). In some embodiments, the NPR1 antibodies or antigen-binding fragments thereof described herein can be combined with one or more corticosteroids (e.g., inhaled corticosteroids, such as fluticasone, budesonide, mometasone, beclomethasone, ciclesonide, or fluticasone furoate, or oral or intravenous corticosteroids, such as prednisone or methylprednisolone), leukotriene modifiers (e.g., montelukast, zafirlukast, or zileuton), bronchodilators (e.g., long-acting beta agonists (such as salmeterol or formoterol), short-acting beta agonists (such as salbutamol or levalbuterol), theophylline, or ipratropium), or combinations thereof (e.g., combinations of fluticasone and salmeterol, budesonide and formoterol, or formoterol and mometasone). In some embodiments, the NPR1 antibodies or antigen-binding fragments thereof described herein can be combined with a β -adrenoceptor antagonist (e.g., timolol, levobunolol, metiprolol, carteolol, or betaxolol), a carbonic anhydrase inhibitor (e.g., acetazolamide, dorzolamide, brinzolamide, or methazolamide), an α 2-adrenoceptor agonist (e.g., brimonidine or aclonidine), a parasympathomimetic (e.g., a choledocinoid such as pilocarpine), a prostaglandin analog (e.g., latanoprost (latanoprost bund), travoprost, bimatoprost, tafluprost), a rho kinase inhibitor (e.g., nervosudil or liparidil), or combinations thereof (e.g., a combination of a rho kinase inhibitor and latanoprost).

Accordingly, the methods of treating a disease associated with loss of NPR1 function described herein may further comprise administering to a subject in need of treatment a second agent.

The term "combination" refers to a fixed combination in one dosage unit form; or a combination administration, wherein the anti-NPR 1 antibody or antigen-binding fragment thereof described herein and the combination partner (e.g., another drug, also referred to as a "therapeutic agent" or a "co-agent" (co-agent), as explained below) can be administered independently at the same time or separately within time intervals, particularly where these time intervals allow the combination partner to exhibit a synergistic (e.g., synergistic) effect. The individual components may be packaged in a kit or separately. One or both components (e.g., powder or liquid) may be reconstituted or diluted to the desired dosage prior to administration. As used herein, the terms "co-administration" or "combined administration" and the like are meant to encompass the administration of selected combination partners to a single subject (e.g., patient) in need thereof, and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration and/or concurrently. As used herein, the term "pharmaceutical combination" means a product resulting from the mixing or combination of more than one therapeutic agent, and includes both fixed and non-fixed combinations of therapeutic agents. The term "fixed combination" means that the therapeutic agents (e.g., the compounds of the present invention and the combination partners) are administered to the patient simultaneously in the form of a single entity or dose. The term "non-fixed combination" means that the therapeutic agents (e.g., a compound of the invention and a combination partner) are administered to a patient as separate entities simultaneously, concurrently or sequentially (without specific time constraints), wherein such administration provides therapeutically effective levels of both compounds in the patient. The latter also applies to mixture therapy, for example the administration of three or more therapeutic agents.

As used herein, the term "pharmaceutical combination" refers to a fixed combination in one dosage unit form, or a non-fixed combination or kit of parts for combined administration, wherein two or more therapeutic agents may be administered independently at the same time or separately within time intervals, in particular wherein these time intervals allow the combination partners to show a cooperative, e.g. synergistic effect.

The term "combination therapy" refers to the administration of two or more therapeutic agents to treat the treated condition or disorder described in this disclosure. Such administration encompasses co-administration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule with a fixed ratio of active ingredients. Alternatively, such administration encompasses co-administration in multiple or separate containers (e.g., tablets, capsules, powders, and liquids) for each active ingredient. The powder and/or liquid may be reconstituted or diluted to the desired dosage prior to administration. Further, such administration also encompasses the use of each type of therapeutic agent in a sequential manner at approximately the same time or at different times. In either case, the treatment regimen will provide the beneficial effects of the drug combination in treating the conditions or disorders described herein.

Pharmaceutical compositions, dosages and methods of administration

Also provided herein are compositions, e.g., pharmaceutical compositions, for treating NPR 1-related diseases. Such compositions include one or more anti-NPR 1 antibodies or antigen-binding fragments thereof as described herein, and may include a pharmaceutically acceptable carrier. Such compositions may further comprise another agent, for example, in accordance with the current standard of care for the disease to be treated.

The pharmaceutical composition typically comprises a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not cause significant irritation to a subject and does not abrogate the biological activity and properties of the administered anti-NPR 1 antibody or antigen-binding fragment and/or any additional therapeutic agent in the composition. Pharmaceutically acceptable carriers may enhance or stabilize the composition, or may be used to facilitate preparation of the composition. Pharmaceutically acceptable carriers can include saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Adjuvants may also be included in any of these formulations. Typically, the pharmaceutical compositions are formulated to be compatible with the intended route of administration. Examples of routes of administration include parenteral (e.g., intravenous, intraarterial, intraperitoneal), oral, intracranial, intrathecal or intranasal (e.g., inhalation), intradermal, subcutaneous or transmucosal administration. In some embodiments, the pharmaceutical composition is formulated to deliver an anti-NPR 1 antibody or antigen-binding fragment thereof to cross the blood-brain barrier. The phrases "physiologically acceptable carrier" and "pharmaceutically acceptable carrier" may be used interchangeably.

As used herein, the term "excipient" refers to an inert substance added to a pharmaceutical composition to further facilitate administration of the active ingredient. Formulations for parenteral administration may, for example, contain excipients such as sterile water or saline, polyalkylene glycols (such as polyethylene glycol), vegetable oils or hydrogenated naphthalenes. Other exemplary excipients include, but are not limited to, calcium bicarbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, ethylene-vinyl acetate copolymer particles, and surfactants, including, for example, polysorbate 20.

The pharmaceutical compositions of the present disclosure can be administered by various methods known in the art. The route and/or mode of administration may vary depending on the desired result. In some embodiments, the administration is intravitreal, intravenous, intramuscular, intraperitoneal, or subcutaneous administration. The pharmaceutically acceptable carrier should be suitable for intravitreal, intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active compound(s) (i.e., the anti-NPR 1 antibody or antigen-binding fragment, and optionally additional therapeutic agents) may be coated in a material to protect the compound(s) from the action of acids and other natural conditions that may inactivate the compound(s).

Typically, a therapeutically effective (effective or effective) dose of the anti-NPR 1 antibody or antigen binding fragment is used in the pharmaceutical compositions of the disclosure. The anti-NPR 1 antibody or antigen-binding fragment can be formulated into a pharmaceutically acceptable dosage form by conventional methods known to those skilled in the art.

Methods of formulating suitable pharmaceutical compositions are known in The art, see, e.g., Remington, The Science and Practice of Pharmacy [ leiminton: pharmaceutical science and practice 21 st edition, 2005; and in Drugs and the Pharmaceutical Sciences a Series of Textbooks and monograms [ Pharmaceutical and Pharmaceutical Sciences: books in the series textbook and monograph (Dekker, new york) series, the contents of each of which are hereby incorporated by reference. For example, a solution or suspension for parenteral, intradermal, or subcutaneous application may include the following components: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers such as acetate, citrate or phosphate; and agents for regulating tonicity, such as sodium chloride or dextrose. The pH can be adjusted with an acid or base, such as hydrochloric acid or sodium hydroxide. The parenteral formulations may be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

The dosing regimen of the anti-NPR 1 antibody and antigen-binding fragment with or without additional therapeutic agents can be adjusted to provide the optimal desired response (e.g., therapeutic response). For example, one or both agents may be administered as a single bolus, several divided doses may be administered within a predetermined time period, or the doses of one or both agents may be proportionally reduced or increased, as indicated by the exigencies of the therapeutic situation. For any particular subject, the particular dosage regimen may be adjusted over time according to the individual need and the professional judgment of the treating clinician. Parenteral compositions can be formulated in dosage unit form for ease of administration and uniformity of dosage. As used herein, a unit dosage form refers to physically discrete units suitable as a single dose for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

An "effective amount" is an amount sufficient to achieve a beneficial or desired result. For example, a therapeutic amount is an amount that achieves a desired therapeutic effect. The amount can be the same or different from a prophylactically effective amount, which is an amount required to prevent onset of a disease or disease symptom. An effective amount may be administered in one or more administrations, applications or administrations. The therapeutically effective amount (i.e., effective dose) of the therapeutic compound depends on the therapeutic compound selected. One skilled in the art (e.g., a physician) will appreciate that certain factors may affect the dosage and timing required to effectively treat a subject, including, but not limited to, the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. In addition, treatment of a subject with a therapeutically effective amount of a therapeutic compound described herein can include a single treatment or a series of treatments.

The dose, toxicity and therapeutic efficacy of a therapeutic compound can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., to determine LD50 (the dose lethal to 50% of the population) and ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED 50. Compounds exhibiting high therapeutic indices are preferred. Although compounds exhibiting toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of the affected tissue so as to minimize potential damage to uninfected cells and thereby reduce side effects.

The dosing regimen of the anti-NPR 1 antibody and antigen-binding fragment (alone or in combination with additional therapeutic agents) can be adjusted to provide the optimal desired response (e.g., therapeutic response). For example, one or both agents may be administered as a single bolus, several divided doses may be administered within a predetermined time period, or the doses of one or both agents may be proportionally reduced or increased, as indicated by the exigencies of the therapeutic situation. For any particular subject, the particular dosage regimen may be adjusted over time according to the individual need and the professional judgment of the treating clinician. Parenteral compositions can be formulated in dosage unit form for ease of administration and uniformity of dosage. As used herein, a unit dosage form refers to physically discrete units suitable as a single dose for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

Dosage values for compositions comprising the anti-NPR 1 antibody or antigen-binding fragment and/or any one or more additional therapeutic agents may be selected based on the unique characteristics of the active compound or compounds and the particular therapeutic effect to be achieved. A physician or veterinarian can start a dose of an antibody of the present disclosure used in a pharmaceutical composition at a level below that required to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved. In general, the effective dosage of a composition of the present disclosure for treating obesity or another disease described herein can vary depending on a number of different factors, including the mode of administration, the target site, the physiological state of the patient, whether the patient is a human or a human animal, other drugs administered, and whether the treatment is prophylactic or therapeutic. The selected dose level may also depend upon a variety of pharmacokinetic factors including the activity of the particular composition of the disclosure or ester, salt or amide thereof employed, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular composition employed, the age, sex, weight, condition, general health and past medical history of the patient being treated. The therapeutic dose can be titrated to optimize safety and efficacy.

Reagent kit

Also provided herein are kits comprising one or more of the compositions provided herein (e.g., an antibody or antigen-binding fragment thereof described in table 2, table 3, or table 4) and instructions for use. The instructions for use may include instructions for diagnosing or treating a disease associated with NPR 1. The kit as provided herein may be used according to any of the methods described herein. One skilled in the art will recognize other suitable uses for the kits provided herein, and will be able to use the kits for such uses. Kits provided herein can also include a mailer (e.g., a postage payment envelope or mailer) that can be used to return a sample for analysis to, for example, a laboratory. The kit may include one or more containers for the sample, or the sample may be in a standard blood collection vial. The kit may further comprise one or more of: informed consent, test application and instructions on how to use the kit in the methods described herein. Also included herein are methods of using such kits. One or more tables (e.g., test application tables) and containers holding samples can be encoded, for example, with a bar code (used to identify the subject providing the sample).

The disclosure is further illustrated by the following examples and claims, which are illustrative and not meant to be further limiting. One skilled in the art will recognize that many methods and materials similar or equivalent to those described herein can be used in the practice of the compositions and methods. Such equivalents are within the scope of the disclosure and claims. The contents of all references, including issued patents and published patent applications, cited throughout this application are hereby incorporated by reference.

Examples

In more detail, the present disclosure provides the following embodiments:

example 1. an isolated antibody or antigen-binding fragment that (i) binds to natriuretic peptide receptor 1(NPR 1); and (ii) is capable of activating NPR1 in the absence of Atrial Natriuretic Peptide (ANP).

Example 2. an isolated anti-NPR 1 antibody or antigen-binding fragment; or an isolated antibody or antigen-binding fragment thereof as described in example 1, which does not bind to and/or does not activate natriuretic peptide receptor 2(NPR2) and/or natriuretic peptide receptor 3(NPR 3).

Example 3. an isolated anti-NPR 1 antibody or antigen-binding fragment; or an isolated antibody or antigen-binding fragment as described in example 1 or example 2, which binds to (a) human NPR 1; and (b) mouse NPR1 and/or rat NPR 1.

Example 4. an isolated anti-NPR 1 antibody or antigen-binding fragment; or an isolated antibody or antigen-binding fragment as described in example 1 or example 2, which binds to (a) human NPR 1; and (b) cynomolgus monkey NPR1 binding.

Example 5 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment thereof of any one of examples 1-4, which is ANP non-competitive.

Example 6. an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment thereof of any one of examples 1, 2, or 4, which is ANP competitive.

Example 7 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen binding fragment of any one of examples 1-5, which is capable of stabilizing the ANP-NPR1 complex.

Example 8 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of embodiments 1-5 or 7, wherein the antibody or antigen-binding fragment thereof binds to an epitope within amino acids 99-133 of SEQ ID NO. 1.

Example 9 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of embodiments 1-5, 7 or 8, wherein the antibody or antigen-binding fragment thereof binds to an epitope comprising at least two amino acid residues within amino acids 99-133 of SEQ ID NO: 1.

Example 10 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of embodiments 1-5 or 7-9, wherein the antibody or antigen-binding fragment thereof binds to an epitope comprising at least 3, 4, 5, 6, 7, or 8 amino acid residues within amino acids 99-133 of SEQ ID NO: 1.

Example 11. an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of embodiments 1-5 or 7-10, wherein the antibody or antigen-binding fragment thereof binds to an epitope within amino acids 99-111 of SEQ ID NO. 1.

Example 12 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of embodiments 1-5 or 7-11, wherein the antibody or antigen-binding fragment thereof binds to an epitope within amino acids 99-103 of SEQ ID NO: 1.

Example 13 an isolated anti-NPR 1 antibody or antigen-binding fragment; or an isolated antibody or antigen-binding fragment as described in any one of examples 1-5 or 7-12, wherein the antibody or antigen-binding fragment thereof binds to an epitope within amino acids 105 and 111 of SEQ ID NO: 1.

Example 14. an isolated anti-NPR 1 antibody or antigen-binding fragment; or an isolated antibody or antigen-binding fragment as described in any one of examples 1-5 or 7-13, wherein the antibody or antigen-binding fragment thereof binds to an epitope comprising at least 2, 3 or 4 amino acid residues within amino acid 105-111 of SEQ ID NO: 1.

Example 15 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any of examples 1-5 or 7-14, wherein the antibody or antigen-binding fragment thereof binds to a conformational epitope of human NPR1, and wherein the conformational epitope comprises at least one amino acid residue of each of (i) amino acids 99-103 of SEQ ID NO:1, (ii) amino acids 105-111 of SEQ ID NO:1, (iii) amino acids 131-134 of SEQ ID NO:1, and further binds to amino acids 375 and/or 378 of SEQ ID NO: 1.

Example 16. an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen binding fragment of any one of embodiments 8-14, wherein the epitope is a conformational epitope, and wherein the conformational epitope additionally comprises at least one amino acid residue selected from the group consisting of amino acids 33, 34, 76, 82, and 104 of SEQ ID No. 1.

Example 17 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen binding fragment of embodiment 15, wherein the conformational epitope additionally comprises at least one amino acid residue selected from the group consisting of amino acids 33, 34, 76, 82, 104, 374, and 375 of SEQ ID No. 1.

Example 18. an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of embodiments 1-5 or 7-17, wherein the antibody or antigen-binding fragment thereof binds to at least

amino acids

82, 102, 103, 105, 106, 109, 132, and 375 of SEQ ID NO: 1.

Example 19 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of embodiments 1-5 or 7-18, wherein the antibody or antigen-binding fragment thereof binds to at least

amino acids

34, 82, 102, 103, 105, 106, 107, 109, 132, 133, 375, and 378 of SEQ ID NO. 1.

Example 20 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of embodiments 1-5 or 7-18, wherein the antibody or antigen-binding fragment thereof binds to at least

amino acids

79, 82, 99, 102, 103, 105, 106, 109, 131, 132, and 375 of SEQ ID NO: 1.

Example 21, an isolated anti-NPR 1 antibody or antigen-binding fragment; or an isolated antibody or antigen-binding fragment as described in any one of examples 1, 2, 4 or 6, wherein the antibody or antigen-binding fragment thereof binds to an epitope within amino acids 188 and 219 of SEQ ID NO: 1.

Example 22. an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of examples 1, 2, 4, 6 or 21, wherein the antibody or antigen-binding fragment thereof binds to an epitope comprising at least 2, 3, 4, 5, 6 or 7 amino acids within amino acids 188 and 219 of SEQ ID NO: 1.

Example 23 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any of examples 1, 2, 4, 6, 21 or 22, wherein the antibody or antigen-binding fragment thereof binds to a conformational epitope within NPR1, and wherein the conformational epitope comprises at least one amino acid residue of each of (i) amino acids 188 and 198 of SEQ ID NO:1, (ii) amino acids 201 and 208 of SEQ ID NO:1, (iii) amino acids 215 and 238 of SEQ ID NO:1, and (iv) amino acids 294 and 297 of SEQ ID NO: 1.

Example 24 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of

embodiments

1, 2, 4, 6, or 21-23, wherein the antibody or antigen-binding fragment thereof binds to at least amino acids 188, 192, 194, 197, 201, 208, and 219 of SEQ ID No. 1.

Example 25 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any one of

embodiments

1, 2, 4, 6, or 21-24, wherein the antibody or antigen-binding fragment thereof binds to at least amino acids 188, 192, 194, 197, 201, 208, 219, and 295 of SEQ ID No. 1.

Example 26 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of embodiments 1-5 or 7-20, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2 and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2 and LCDR3), and wherein the antibody or antigen-binding fragment comprises CDRs from one of the ANP non-competitive groups set forth in table 3 or table 4.

Example 27 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of

embodiments

1, 2, 4, 6, or 21-25, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), and wherein the antibody or antigen-binding fragment comprises CDRs of one of the ANP competitive groups set forth in table 3 or table 4.

Example 28. an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of embodiments 1-5, 7-20, or 26, wherein the antibody or antigen-binding fragment is WW01_ LALA, WW03_ LALA, XX01_ LALA, XX01_ DAPA, XX01_ N30S _ DAPA, XX03_ LALA, XX04_ LALA, XX06_ LALA, XX06_ DAPA, XX07_ LALA, XX08_ LALA, XX08_ DAPA, XX08_ N30S _ DAPA, XX08_ N30Q _ DAPA, XX09_ LALA, XX11_ LALA, XX12_ LALA, XX13_ LALA, XX14_ LALA, 14_ laxx, 14_ laja, 14_ laxx, 14_ laja, 36yja, 14_ laja, 14, 36laja _ laja, 14, 36yja _ laja, 14, 36laja _ laja, 14, 36laja _ laja, 14, 36yay _ laja, 14, 36laja _ laja, 14, 36laja _ laja _ la.

Example 29 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of

embodiments

1, 2, 4, 6, 21-25, or 27, wherein the antibody or antigen-binding fragment is WW05_ LALA, WW06_ LALA, YY05_ LALA, YY06_ LALA, YY07_ LALA, ZZ05_ LALA, ZZ14_ LALA, and ZZ16_ LALA.

Example 30 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of embodiments 1-5, 7-20, 26, or 28, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), and wherein:

(a) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 28; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆YADSVKG (SEQ ID NO:429) or consists of the amino acid sequence shown in YADSVKG (SEQ ID NO:429), wherein X₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X₆Is Y or F; HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30; LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41; LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 42; and LCDR3 contains, for example, Y₁QY₂Y₃Y₄Y₅An amino acid sequence shown by PRT (SEQ ID NO:430) or consists of the amino acid sequence; wherein Y is₁Is M or Q, Y₂Is S, E, T or I, Y₃Is Y or W, Y₄Is E, V, R, A, T or M, and Y₅Is K, V, R or A;

(II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 31; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆Ammonia as shown in YADSVKG (SEQ ID NO:429)An amino acid sequence or consisting thereof, wherein X₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X₆Is Y or F; the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 42, and the LCDR3 comprises or consists of Y ₁QY₂Y₃Y₄Y₅An amino acid sequence shown by PRT (SEQ ID NO:430) or consists of the amino acid sequence; wherein Y is₁Is M or Q, Y₂Is S, E, T or I, Y₃Is Y or W, Y₄Is E, V, R, A, T or M, and Y₅Is K, V, R or A;

(III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:32, HCDR2 comprises₁SX₂GX₃Y (SEQ ID NO:431) or an amino acid sequence consisting thereof, wherein X₁Is S or E, X₂Is D or K, or X₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 44, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 45, and the LCDR3 comprises or consists of the amino acid sequence shown as Y₁Y₂Y₃Y₄PR (SEQ ID NO:432) or consists of the amino acid sequence thereof; wherein Y is₁Is S, E, T or I, Y₂Is Y or W, Y₃Is E, V, R, A, T or M, and Y₄Is K, V, R or A; or

(IV) HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:34 and HCDR2 comprises or consists of the amino acid sequence shown in IX₁SX₂GX₃YX₄(SEQ ID NO:433) or consists of the amino acid sequence shown in (wherein X is₁Is S or E, X₂Is D or K, X₃Is S or N, and X₄Is I or T, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:36, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:47, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and LCDR3 comprises or consists of Y ₁QY₂Y₃Y₄Y₅An amino acid sequence shown by PRT (SEQ ID NO:430) or consists of the amino acid sequence; wherein Y is₁Is M or Q, Y₂Is S, E, T or I, Y₃Is Y or W, Y₄Is E, V, R, A, T or M, and Y₅Is K, V, R or A;

(b) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 28; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆YADSVKG (SEQ ID NO:429) or consists of the amino acid sequence shown in YADSVKG (SEQ ID NO:429), wherein X₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X₆Is Y or F; HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30; LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41; LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 42; and LCDR3 comprises e.g. QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A;

(II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 31; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆YADSVKG (SEQ ID NO:429) or consists of the amino acid sequence shown in YADSVKG (SEQ ID NO:429), wherein X₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X₆Is Y or F; the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 42, and the LCDR3 comprises or consists of the amino acid sequence shown as QQY ₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A;

(III) HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:32, HCDR2Containing as X₁SX₂GX₃Y (SEQ ID NO:431) or an amino acid sequence consisting thereof, wherein X₁Is S or E, X₂Is D or K, or X₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 44, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 45, and the LCDR3 comprises or consists of the amino acid sequence shown as Y₁WY₂Y₃PR (SEQ ID NO:435) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; or

(IV) HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:34 and HCDR2 comprises or consists of the amino acid sequence shown in IX₁SX₂GX₃YX₄(SEQ ID NO:433) or consists of the amino acid sequence shown in (wherein X is₁Is S or E, X₂Is D or K, X₃Is S or N, and X₄Is I or T, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:36, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:47, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and LCDR3 comprises or consists of the amino acid sequence shown as QQY ₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A;

(c) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 28; HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 119; HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30; LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41; LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 42; and LCDR3 comprises e.g. QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A;

(II) HCDR1 comprises ammonia as shown in SEQ ID NO:31An amino acid sequence or consisting thereof; HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 119; the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 42, and the LCDR3 comprises or consists of the amino acid sequence shown as QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is ₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A;

(III) the HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:32, the HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:120, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:44, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and the LCDR3 comprises or consists of the amino acid sequence shown as Y₁WY₂Y₃PR (SEQ ID NO:435) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; or

(IV) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:34, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:121, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:36, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:47, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and LCDR3 comprises or consists of the amino acid sequence shown as QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y ₃Is K, V, R or A;

(d) (I) HCDR1 comprises e.g. GFTFX₁An amino acid sequence shown by THYIH (SEQ ID NO:436) or consists of the same, wherein X₁Is N, S or Q, HCDR2 comprises e.g. SIY₁Y₂Y₃GY₄Y₅TY₆YADSVKG (SEQ ID NO:437) or an amino acid sequence consisting of the same, wherein Y₁Is S or G, Y₂Is S or G, Y₃Is S or Q, Y₄Is S, Q or G, Y₅Is S, N or M, and Y₆Is Y or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 17, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 18, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19;

(II) HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:7 and HCDR2 comprises or consists of SIY₁Y₂Y₃GY₄Y₅TY₆YADSVKG (SEQ ID NO:437) or an amino acid sequence consisting of the same, wherein Y₁Is S or G, Y₂Is S or G, Y₃Is S or Q, Y₄Is S, Q or G, Y₅Is S, N or M, and Y₆Is Y or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 17, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 18, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19;

(III) HCDR1 comprises e.g. GFTFX₁TH (SEQ ID NO:438) or an amino acid sequence consisting thereof, wherein X₁Is N, S or Q, HCDR2 comprises e.g. Y₁Y₂Y₃GY₄Y₅(SEQ ID NO:439) or an amino acid sequence consisting thereof, wherein Y₁Is S or G, Y₂Is S or G, Y₃Is S or Q, Y₄Is S, Q or G, and Y₅S, N or M, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 20, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 21, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 22; or

(IV) HCDR1 contains e.g. GFTFX₁An amino acid sequence represented by THY (SEQ ID NO:440) or consisting thereof, wherein X₁Is N, S or Q, HCDR2 comprises, for example, IY₁Y₂Y₃GY₄Y₅T (SEQ ID NO:441) or an amino acid sequence consisting thereof, wherein Y₁Is S or G, Y₂Is S or G, Y₃Is S or Q, Y₄Is S, Q or G, and Y₅S, N or M, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 12, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 23, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 21, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19;

(e) (I) HCDR1 comprises e.g. GFTFX₁An amino acid sequence shown by THYIH (SEQ ID NO:436) or consists of the same, wherein X₁Is N, S or Q, HCDR2 comprises e.g. SISY₁SGY₂Y₃TYYADSVKG (SEQ ID NO:442), wherein Y is₁Is S or G, Y₂Is S or Q, and Y₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 17, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 18, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19;

(II) HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:7 and HCDR2 comprises a sequence as SISY₁SGY₂Y₃TYYADSVKG (SEQ ID NO:442), wherein Y is₁Is S or G, Y₂Is S or Q, and Y₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 17, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 18, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19;

(III) HCDR1 comprises e.g. GFTFX₁TH (SEQ ID NO:438) or an amino acid sequence consisting thereof, wherein X ₁Is N, S or Q, HCDR2 comprises e.g. SY₁SGY₂Y₃(SEQ ID NO:443) or an amino acid sequence consisting of the same, wherein Y is₁Is S or G, Y₂Is S or Q, and Y₃Is S or N, HCDR3 comprisesThe amino acid sequence shown as SEQ ID NO. 6 or consists thereof, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 20, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 21, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 22; or

(IV) HCDR1 contains e.g. GFTFX₁An amino acid sequence represented by THY (SEQ ID NO:440) or consisting thereof, wherein X₁Is N, S or Q, HCDR2 comprises, for example, ISY₁SGY₂Y₃T (SEQ ID NO:444) or an amino acid sequence consisting thereof, wherein Y₁Is S or G, Y₂Is S or Q, and Y₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 12, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 23, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 21, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19;

(f) (I) HCDR1 contains e.g. GFX₁FSX₂YX₃X₄X₅(SEQ ID NO:445) or a sequence consisting of the amino acid sequence shown in (SEQ ID NO:445), wherein X₁Is S or T, X₂Is S, K or R, X₃Is W or Y, X ₄Is I or L, and X₅Is S or N, HCDR2 comprises, e.g., Y₁IY₂QY₃Y₄Y₅EY₆Y₇YVESVKG (SEQ ID NO:446) or a sequence consisting of the same, wherein Y is₁Is S or N, Y₂Is K or H, Y₃Is S, Q or H, Y₄Is G or A, Y₅Is S, H or L, Y₆Is T or K, and Y₇Y, K or R, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(II) HCDR1 contains as X₁YX₂X₃X₄(SEQ ID NO:447) or consists of an amino acid sequence as shown in, wherein X₁Is S, K or R, X₂Is W or Y, X₃Is I or L, and X₄Is S or N, HCDR2 comprises, e.g., Y₁IY₂QY₃Y₄Y₅EY₆Y₇YVESVKG (SEQ ID NO:446) or a sequence consisting of the same, wherein Y is₁Is S or N, Y₂Is K or H, Y₃Is S, Q or H, Y₄Is G or A, Y₅Is S, H or L, Y₆Is T or K, and Y₇Y, K or R, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(III) HCDR1 contains, for example, GFX₁FSX₂Y (SEQ ID NO:448) or an amino acid sequence consisting of the same, wherein X₁Is S or T, and X₂Is S, K or R, HCDR2 comprises e.g. Y₁QY₂Y₃Y₄E (SEQ ID NO:449) or an amino acid sequence consisting of the same, wherein Y₁Is K or H, Y₂Is S, Q or H, Y₃Is G or A, and Y₄S, H or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or

(IV) HCDR1 contains, for example, GFX₁FSX₂YX₃(SEQ ID NO:450) or consists of an amino acid sequence shown in (SEQ ID NO:450), wherein X₁Is S or T, X₂Is S, K or R, and X₃Is W or Y, HCDR2 comprises, e.g., IY₁QY₂Y₃Y₄EY₅(SEQ ID NO:451) or consists of an amino acid sequence shown in (SEQ ID NO:451), wherein Y₁Is K or H, Y₂Is S, Q or H, Y₃Is G or A, Y₄Is S, H or L, and Y₅Is T or K, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:232, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:243, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241 and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(g) (I) HCDR1 contains e.g. GFX₁FSX₂YX₃X₄X₅(SEQ ID NO:445) or a sequence consisting of the amino acid sequence shown in (SEQ ID NO:445), wherein X₁Is S or T, X₂Is S, K or R, X₃Is W or Y, X₄Is I or L, and X₅Is S or N, HCDR2 comprises e.g. SIHQY₁Y₂Y₃EY₄Y₅YVESVKG (SEQ ID NO:453) or consist of the amino acid sequence shown in, wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, Y₄Is T or K, and Y₅Is K or R, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(II) HCDR1 contains as X₁YX₂X₃X₄(SEQ ID NO:447) or consists of an amino acid sequence as shown in, wherein X₁Is S, K or R, X₂Is W or Y, X₃Is I or L, and X₄Is S or N, HCDR2 comprises e.g. SIHQY₁Y₂Y₃EY₄Y₅YVESVKG (SEQ ID NO:453) or consist of the amino acid sequence shown in, wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, Y₄Is T or K, and Y₅Is K or R, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(III) HCDR1 contains, for example, GFX₁FSX₂Y (SEQ ID NO:448) or an amino acid sequence consisting of the same, wherein X₁Is S or T, and X₂Is S, K or R, HCDR2 comprises e.g. HQY₁Y₂Y₃E (SEQ ID NO:456) or an amino acid sequence consisting of the same, wherein Y₁Is Q or H, Y₂Is G or A, and Y₃Is H or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or

(IV) HCDR1 contains, for example, GFX₁FSX₂YX₃(SEQ ID NO:450) or consists of an amino acid sequence shown in (SEQ ID NO:450), wherein X₁Is S or T, X₂Is S, K or R, and X₃Is W or Y, HCDR2 includes, for example, IHQY₁Y₂Y₃EY₄(SEQ ID NO:458) or consists of an amino acid sequence shown in (SEQ ID NO:458) wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, and Y₄Is T or K, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:232, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:243, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(h) (I) HCDR1 comprises, for example, GFTFSX₁YX₂IX₃(SEQ ID NO:452) or consists of an amino acid sequence as shown in (SEQ ID NO:452), wherein X₁Is S or R, X₂Is W or Y, and X₃Is S or N, HCDR2 comprises, e.g., Y₁IY₂QY₃Y₄Y₅EY₆Y₇YVESVKG (SEQ ID NO:446) or a sequence consisting of the same, wherein Y is₁Is S or N, Y₂Is K or H, Y₃Is S, Q or H, Y₄Is G or A, Y₅Is S, H or L, Y₆Is T or K, and Y₇Y, K or R, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:239Forming;

(II) HCDR1 contains as X₁YX₂IX₃(SEQ ID NO:454) or consists of an amino acid sequence shown in (SEQ ID NO:454), wherein X₁Is S or R, X₂Is W or Y, and X₃Is S or N, HCDR2 comprises, e.g., Y₁IY₂QY₃Y₄Y₅EY₆Y₇YVESVKG (SEQ ID NO:446) or a sequence consisting of the same, wherein Y is₁Is S or N, Y₂Is K or H, Y₃Is S, Q or H, Y₄Is G or A, Y₅Is S, H or L, Y₆Is T or K, and Y₇Y, K or R, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(III) HCDR1 comprises, for example, GFTFSX₁Y (SEQ ID NO:455) or an amino acid sequence consisting thereof, wherein X₁Is S or R, HCDR2 comprises e.g. Y1QY₂Y₃Y₄E (SEQ ID NO:449) or an amino acid sequence consisting of the same, wherein Y₁Is K or H, Y₂Is S, Q or H, Y₃Is G or A, and Y₄S, H or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or

(IV) HCDR1 comprises, for example, GFTFSX₁YX₂(SEQ ID NO:457) or an amino acid sequence consisting of the amino acid sequence, wherein X₁Is S or R, and X₂Is W or Y, HCDR2 comprises, e.g., IY₁QY₂Y₃Y₄EY₅(SEQ ID NO:451) or consists of an amino acid sequence shown in (SEQ ID NO:451), wherein Y₁Is K or H, Y₂Is S, Q or H, Y₃Is G or A, Y₄Is S, H or L, and Y₅Is T or K, HCDR3 comprises the amino acid sequence shown in SEQ ID NO:232Or consists thereof, the LCDR1 comprises or consists of the amino acid sequence as shown in SEQ ID No. 243, the LCDR2 comprises or consists of the amino acid sequence as shown in SEQ ID No. 241 and the LCDR3 comprises or consists of the amino acid sequence as shown in SEQ ID No. 239; or

(i) (I) HCDR1 comprises, for example, GFTFSX₁YX₂IX₃(SEQ ID NO:452) or consists of an amino acid sequence as shown in (SEQ ID NO:452), wherein X₁Is S or R, X₂Is W or Y, and X₃Is S or N, HCDR2 comprises e.g. SIHQY₁Y₂Y₃EY₄Y₅YVESVKG (SEQ ID NO:453) or consist of the amino acid sequence shown in, wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, Y₄Is T or K, and Y₅Is K or R, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(II) HCDR1 contains as X₁YX₂IX₃(SEQ ID NO:454) or consists of an amino acid sequence shown in (SEQ ID NO:454), wherein X₁Is S or R, X₂Is W or Y, and X₃Is S or N, HCDR2 comprises e.g. SIHQY₁Y₂Y₃EY₄Y₅YVESVKG (SEQ ID NO:453) or consist of the amino acid sequence shown in, wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, Y₄Is T or K, and Y₅Is K or R, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:237, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:238, and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(III) HCDR1 comprises, for example, GFTFSX₁Y (SEQ ID NO:455) or an amino acid sequence consisting thereof, wherein X₁Is S or R, HCDR2 comprises e.g. HQY1Y₂Y₃E (SEQ ID NO:456) or consists ofWherein Y is₁Is Q or H, Y₂Is G or A, and Y₃Is H or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or

(IV) HCDR1 comprises, for example, GFTFSX₁YX₂(SEQ ID NO:457) or an amino acid sequence consisting of the amino acid sequence, wherein X₁Is S or R, and X₂Is W or Y, HCDR2 includes, for example, IHQY₁Y₂Y₃EY₄(SEQ ID NO:458) or consists of an amino acid sequence shown in (SEQ ID NO:458) wherein Y is₁Is Q or H, Y₂Is G or A, Y₃Is H or L, and Y₄Is T or K, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:232, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:243, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239.

Example 31. an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of embodiments 1, 2, 4, 6, 21-25, 27, or 29, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), and wherein:

(a) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:310, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of GX₁X₂X₃GX₄LGFDH (SEQ ID NO:459) or consists of the amino acid sequence shown in the specification, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:320, LCDR2 comprises, for example, GNSNRPY₁(SEQ ID NO:460) or consists of an amino acid sequence shown in (SEQ ID NO:460) wherein Y₁Is S or N, and LCDR3 comprises, for example, QSYZ₁Z₂Z₃Z₄Z₅Z₆Z₇V (SEQ ID NO:461) wherein Z is₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V;

(II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:229, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of GX ₁X₂X₃GX₄LGFDH (SEQ ID NO:459) or consists of the amino acid sequence shown in the specification, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:320, LCDR2 comprises, for example, GNSNRPY₁(SEQ ID NO:460) or consists of an amino acid sequence shown in (SEQ ID NO:460) wherein Y₁Is S or N, and LCDR3 comprises, for example, QSYZ₁Z₂Z₃Z₄Z₅Z₆Z₇V (SEQ ID NO:461) wherein Z is₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V;

(III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:80, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:313, HCDR3 comprises or consists of GX₁X₂X₃GX₄LGFDH (SEQ ID NO:459) or consists of the amino acid sequence shown in the specification, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:323, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, and LCDR3 comprises or consists of the amino acid sequence shown as YZ₁Z₂Z₃Z₄Z₅Z₆Z₇(SEQ ID NO:462) or an amino acid sequence consisting of the amino acid sequence shown in (SEQ ID NO:462), wherein Z₁Is Y,D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z ₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V; or

(IV) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:82, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:314, HCDR3 comprises or consists of ARGX₁X₂X₃GX₄LGFDH (SEQ ID NO:463) or an amino acid sequence consisting of₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:326, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, and LCDR3 comprises or consists of the amino acid sequence shown as QSYZ₁Z₂Z₃Z₄Z₅Z₆Z₇V (SEQ ID NO:461) wherein Z is₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V;

(b) (I) HCDR1 comprises e.g. GFTFX₁X₂YAX₃X₄(SEQ ID NO:464) or consists of an amino acid sequence as shown in (SEQ ID NO:464), wherein X₁Is S or G, X₂Is S or T, X₃Is I or M, and X₄Is S or T, HCDR2 comprises e.g. Y₁ISY₂Y₃GY₄Y₅Y₆Y₇YAY₈An amino acid sequence as shown in SVKG (SEQ ID NO:465) or consists thereof, wherein Y₁Is A or S, Y₂Is A, S or G, Y₃Is S or H, Y₄Is G or Y, Y₅Is S or Y, Y₆Is T or A, Y₇Is Y, R or N, and Y ₈Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339;

(II) HCDR1 contains as X₁YAX₂X₃(SEQ ID NO:466) or consists of an amino acid sequence shown in (SEQ ID NO:466) wherein X₁Is S or T, X₂Is I or M, and X₃Is S or T, HCDR2 comprises e.g. Y₁ISY₂Y₃GY₄Y₅Y₆Y₇YAY₈An amino acid sequence as shown in SVKG (SEQ ID NO:465) or consists thereof, wherein Y₁Is A or S, Y₂Is A, S or G, Y₃Is S or H, Y₄Is G or Y, Y₅Is S or Y, Y₆Is T or A, Y₇Is Y, R or N, and Y₈Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339;

(III) HCDR1 comprises e.g. GFTFX₁X₂Y (SEQ ID NO:467), wherein X is₁Is S or G, and X₂Is S or T, HCDR2 comprises e.g. SY₁Y₂GY₃Y₄(SEQ ID NO:468) or a sequence consisting of the amino acids shown in (SEQ ID NO: Y) ₁Is A, S or G, Y₂Is S or H, Y₃Is G or Y, and Y₄Is S or Y, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:340, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 342; or

(IV) HCDR1 contains e.g. GFTFX₁X₂YA (SEQ ID NO:469) or an amino acid sequence consisting of the same, wherein X₁Is S or G, and X₂Is S or T, HCDR2 comprises, e.g., ISY₁Y₂GY₃Y₄T (SEQ ID NO:470) or an amino acid sequence consisting thereof, wherein Y₁Is S or G, Y₂Is S or H, Y₃Is G or Y, and Y₄Is S or Y, HCDR3 comprises an amino acid sequence as set forth in SEQ ID332, LCDR1 comprising or consisting of the amino acid sequence shown as SEQ ID NO:343, LCDR2 comprising or consisting of the amino acid sequence shown as SEQ ID NO:341 and LCDR3 comprising or consisting of the amino acid sequence shown as SEQ ID NO: 339; or

(c) (I) HCDR1 comprises e.g. GFTFX₁X₂YAX₃X₄(SEQ ID NO:464) or consists of an amino acid sequence as shown in (SEQ ID NO:464), wherein X₁Is S or G, X₂Is S or T, X₃Is I or M, and X₄Is S or T, HCDR2 comprises e.g. SISY ₁Y₂GYYY₃Y₄YAY₅An amino acid sequence as shown in SVKG (SEQ ID NO:471) or consists thereof, wherein Y₁Is A or S, Y₂Is S or H, Y₃Is T or A, Y₄Is R or N, and Y₅Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339;

(II) HCDR1 contains as X₁YAX₂X₃(SEQ ID NO:466) or consists of an amino acid sequence shown in (SEQ ID NO:466) wherein X₁Is S or T, X₂Is I or M, and X₃Is S or T, HCDR2 comprises e.g. SISY₁Y₂GYYY₃Y₄YAY₅An amino acid sequence as shown in SVKG (SEQ ID NO:471) or consists thereof, wherein Y₁Is A or S, Y₂Is S or H, Y₃Is T or A, Y₄Is R or N, and Y₅Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339;

(III) HCDR1 comprises e.g. GFTFX₁X₂Y (SEQ ID NO:467), wherein X is ₁Is S or G, and X₂Is S or T, HCDR2 comprises e.g. SY₁Y₂GYY (SEQ ID NO:472) or an amino acid sequence consisting of the same, wherein Y is₁Is A or S, and Y₂Is S or H, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:340, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 342; or

(IV) HCDR1 contains e.g. GFTFX₁X₂YA (SEQ ID NO:469) or an amino acid sequence consisting of the same, wherein X₁Is S or G, and X₂Is S or T, HCDR2 comprises, e.g., ISY₁Y₂G (SEQ ID NO:473) or an amino acid sequence consisting thereof, wherein Y₁Is A, S or G, and Y₂Is S or H, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:332, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:343, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339.

Example 32 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of embodiments 1-5, 7-20, 26, 28, or 30, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), and wherein:

(a) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 28, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 29, 119 and 190, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 30, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 41, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 42, and LCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 43, 126, 134, 145, 172, 178 and 184; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 31, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 29, 119 and 190, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 30, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 41, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 42, and LCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 43, 126, 134, 145, 172, 178 and 184; (III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:32, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID NOs 33, 120 and 191, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:30, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:44, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:45, and LCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID NOs 46, 127, 135, 146, 173, 179 and 185; or (IV) the HCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 34, the HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NO. 35, 121 and 192, the HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 36, the LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 47, the LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 45, and the LCDR3 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NO. 43, 126, 134, 145, 172, 178 and 184;

(b) (I) HCDR1 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 4, 112 and 165, HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 5, 100 and 151, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 6, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID No. 17, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID No. 18, and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 19; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 7, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID nos. 5, 100 and 151, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 17, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 18, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 19; (III) HCDR1 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 8, 113 and 166, HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 9, 101 and 152, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 6, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID No. 20, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID No. 21 and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 22; or (IV) the HCDR1 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO. 10, 114 and 167, the HCDR2 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO. 11, 102 and 153, the HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 12, the LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 23, the LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 21, and the LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO. 19; or

(c) (I) HCDR1 comprising or consisting of the amino acid sequence shown as any one of SEQ ID NOs 226, 367 and 378, HCDR2 comprising or consisting of the amino acid sequence shown as any one of SEQ ID NOs 227, 368 and 379, HCDR3 comprising or consisting of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprising or consisting of the amino acid sequence shown as SEQ ID NO:237, LCDR2 comprising or consisting of the amino acid sequence shown as SEQ ID NO:238, and LCDR3 comprising or consisting of the amino acid sequence shown as SEQ ID NO: 239; (II) HCDR1 comprises or consists of an amino acid sequence as shown in any one of SEQ ID nos. 229, 369 and 380, HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID nos. 227, 368 and 379, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 228, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID No. 237, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID No. 238, and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 239; (III) HCDR1 comprises or consists of the amino acid sequence as shown in any one of SEQ ID nos. 32, 370 and 381, HCDR2 comprises or consists of the amino acid sequence as shown in any one of SEQ ID nos. 230, 371 and 382, HCDR3 comprises or consists of the amino acid sequence as shown in SEQ ID No. 228, LCDR1 comprises or consists of the amino acid sequence as shown in SEQ ID No. 240, LCDR2 comprises or consists of the amino acid sequence as shown in SEQ ID No. 241, and LCDR3 comprises or consists of the amino acid sequence as shown in SEQ ID No. 242; or (IV) HCDR1 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO:34, 372 and 383, HCDR2 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO:231, 373 and 384, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO:232, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO:243, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID NO:241 and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO: 239.

Example 33 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of embodiments 1, 2, 4, 6, 21-25, 27, 29, or 31, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), and wherein:

(a) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:310, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 312 and 348, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:320, LCDR2 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 321 and 354, and LCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 322, 355 and 361; (II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:229, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 312 and 348, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:320, LCDR2 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 321 and 354, and LCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs 322, 355 and 361; (III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:80, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:313, HCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs: 312 and 348, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:323, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, and LCDR3 comprises or consists of the amino acid sequence shown as any of SEQ ID NOs: 325, 356 and 362; or (IV) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:82, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:314, HCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID NO:315 and 349, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:326, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, and LCDR3 comprises or consists of the amino acid sequence shown as any one of SEQ ID NO:322, 355 and 361; or

(b) (I) HCDR1 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 270 and 407, HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs 271, 389 and 408, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO 331, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID No. 337, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID No. 338 and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 339; (II) HCDR1 comprises or consists of the amino acid sequence shown as any one of SEQ ID NOs 273 and 409, HCDR2 comprises or consists of the amino acid sequence shown as any one of SEQ ID NOs 271, 389 and 408, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO 331, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO 337, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO 338, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO 339; (III) HCDR1 comprises or consists of an amino acid sequence as shown in any one of SEQ ID nos. 32 and 410, HCDR2 comprises or consists of an amino acid sequence as shown in any one of SEQ ID nos. 274, 390 and 411, HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 331, LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID No. 340, LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID No. 341, and LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID No. 342; or (IV) the HCDR1 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO:275 and 412, the HCDR2 comprises or consists of an amino acid sequence as shown in any of SEQ ID NO:276, 391 and 413, the HCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO:332, the LCDR1 comprises or consists of an amino acid sequence as shown in SEQ ID NO:343, the LCDR2 comprises or consists of an amino acid sequence as shown in SEQ ID NO:341, and the LCDR3 comprises or consists of an amino acid sequence as shown in SEQ ID NO: 339.

Example 34 an isolated anti-NPR 1 antibody or antigen binding fragment; or the antibody or antigen-binding fragment of any one of embodiments 1-5, 7-20, 26, 28, 30, or 32, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from the group consisting of seq id no:

(a) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 134(LCDR 3);

(b) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 126(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:126(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 126(LCDR 3);

(c) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 145(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:145(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:146(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 145(LCDR 3);

(d) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 172(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 172(LCDR 3);

(e) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 178(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:178(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 178(LCDR 3);

(f) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 184(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:184(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:185(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 184(LCDR 3);

(g) (I) SEQ ID NO 4(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 101(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3);

(h) (I) SEQ ID NO:112(HCDR1), SEQ ID NO:100(HCDR2), SEQ ID NO:6(HCDR3), SEQ ID NO:17(LCDR1), SEQ ID NO:18(LCDR2) and SEQ ID NO:19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 100(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 113(HCDR1), SEQ ID NO 101(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 114(HCDR1), SEQ ID NO 102(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3);

(i) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 119(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 43(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:119(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:43(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:120(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:46(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 121(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 43(LCDR 3);

(j) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 126(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:126(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:127(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 126(LCDR 3);

(k) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 134(LCDR 3);

(l) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 145(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:145(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:146(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 145(LCDR 3);

(m) (I) SEQ ID NO 4(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3);

(n) (I) SEQ ID NO:112(HCDR1), SEQ ID NO:151(HCDR2), SEQ ID NO:6(HCDR3), SEQ ID NO:17(LCDR1), SEQ ID NO:18(LCDR2) and SEQ ID NO:19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 113(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 114(HCDR1), SEQ ID NO 153(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3);

(o) (I) SEQ ID NO:165(HCDR1), SEQ ID NO:151(HCDR2), SEQ ID NO:6(HCDR3), SEQ ID NO:17(LCDR1), SEQ ID NO:18(LCDR2) and SEQ ID NO:19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 151(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 166(HCDR1), SEQ ID NO 152(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO:167(HCDR1), SEQ ID NO:153(HCDR2), SEQ ID NO:12(HCDR3), SEQ ID NO:23(LCDR1), SEQ ID NO:21(LCDR2) and SEQ ID NO:19(LCDR 3);

(p) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 172(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 172(LCDR 3);

(q) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 178(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:178(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:179(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 178(LCDR 3);

(r) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 184(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:184(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:185(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 184(LCDR 3);

(s) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 190(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 134(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:190(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:134(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:191(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:135(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 192(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 134(LCDR 3);

(t) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 190(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 172(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:190(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:172(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:191(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2) and SEQ ID NO:173(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 192(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 172(LCDR 3);

(u) (I) SEQ ID NO 4(HCDR1), SEQ ID NO 5(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (II) SEQ ID NO 7(HCDR1), SEQ ID NO 5(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 17(LCDR1), SEQ ID NO 18(LCDR2) and SEQ ID NO 19(LCDR 3); (III) SEQ ID NO 8(HCDR1), SEQ ID NO 9(HCDR2), SEQ ID NO 6(HCDR3), SEQ ID NO 20(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 22(LCDR 3); or (IV) SEQ ID NO 10(HCDR1), SEQ ID NO 11(HCDR2), SEQ ID NO 12(HCDR3), SEQ ID NO 23(LCDR1), SEQ ID NO 21(LCDR2) and SEQ ID NO 19(LCDR 3);

(v) (I) SEQ ID NO 28(HCDR1), SEQ ID NO 29(HCDR2), SEQ ID NO 30(HCDR3), SEQ ID NO 41(LCDR1), SEQ ID NO 42(LCDR2) and SEQ ID NO 43(LCDR 3); (II) SEQ ID NO:31(HCDR1), SEQ ID NO:29(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:41(LCDR1), SEQ ID NO:42(LCDR2) and SEQ ID NO:43(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:33(HCDR2), SEQ ID NO:30(HCDR3), SEQ ID NO:44(LCDR1), SEQ ID NO:45(LCDR2), and SEQ ID NO:46(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 35(HCDR2), SEQ ID NO 36(HCDR3), SEQ ID NO 47(LCDR1), SEQ ID NO 45(LCDR2) and SEQ ID NO 43(LCDR 3);

(w) (I) SEQ ID NO:367(HCDR1), SEQ ID NO:368(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:239(LCDR 3); (II) SEQ ID NO 369(HCDR1), SEQ ID NO 368(HCDR2), SEQ ID NO 228(HCDR3), SEQ ID NO 237(LCDR1), SEQ ID NO 238(LCDR2) and SEQ ID NO 239(LCDR 3); (III) SEQ ID NO:370(HCDR1), SEQ ID NO:371(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3); or (IV) SEQ ID NO:372(HCDR1), SEQ ID NO:373(HCDR2), SEQ ID NO:232(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:239(LCDR 3);

(x) (I) SEQ ID NO:378(HCDR1), SEQ ID NO:379(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2) and SEQ ID NO:239(LCDR 3); (II) SEQ ID NO:380(HCDR1), SEQ ID NO:379(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2) and SEQ ID NO:239(LCDR 3); (III) SEQ ID NO:381(HCDR1), SEQ ID NO:382(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3); or (IV) SEQ ID NO 383(HCDR1), SEQ ID NO 384(HCDR2), SEQ ID NO 232(HCDR3), SEQ ID NO 243(LCDR1), SEQ ID NO 241(LCDR2), and SEQ ID NO 239(LCDR 3);

(y) (I) SEQ ID NO:226(HCDR1), SEQ ID NO:227(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2) and SEQ ID NO:239(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:227(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2) and SEQ ID NO:239(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:230(HCDR2), SEQ ID NO:228(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:242(LCDR 3); or (IV) SEQ ID NO 34(HCDR1), SEQ ID NO 231(HCDR2), SEQ ID NO 232(HCDR3), SEQ ID NO 243(LCDR1), SEQ ID NO 241(LCDR2) and SEQ ID NO 239(LCDR 3);

(z) (I) SEQ ID NO:270(HCDR1), SEQ ID NO:271(HCDR2), SEQ ID NO:272(HCDR3), SEQ ID NO:282(LCDR1), SEQ ID NO:261(LCDR2), and SEQ ID NO:283(LCDR 3); (II) SEQ ID NO:273(HCDR1), SEQ ID NO:271(HCDR2), SEQ ID NO:272(HCDR3), SEQ ID NO:282(LCDR1), SEQ ID NO:261(LCDR2), and SEQ ID NO:283(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:274(HCDR2), SEQ ID NO:272(HCDR3), SEQ ID NO:284(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:285(LCDR 3); or (IV) SEQ ID NO 275(HCDR1), SEQ ID NO 276(HCDR2), SEQ ID NO 277(HCDR3), SEQ ID NO 286(LCDR1), SEQ ID NO 241(LCDR2), and SEQ ID NO 283(LCDR 3); or

(aa) (I) SEQ ID NO:291(HCDR1), SEQ ID NO:292(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2), and SEQ ID NO:304(LCDR 3); (II) SEQ ID NO:294(HCDR1), SEQ ID NO:292(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:237(LCDR1), SEQ ID NO:238(LCDR2) and SEQ ID NO:304(LCDR 3); (III) SEQ ID NO:295(HCDR1), SEQ ID NO:296(HCDR2), SEQ ID NO:293(HCDR3), SEQ ID NO:240(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:305(LCDR 3); or (IV) SEQ ID NO:297(HCDR1), SEQ ID NO:298(HCDR2), SEQ ID NO:299(HCDR3), SEQ ID NO:243(LCDR1), SEQ ID NO:241(LCDR2), and SEQ ID NO:304(LCDR 3).

Example 35. an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of

embodiments

1, 2, 4, 6, 21-25, 27, 29, 31, or 33, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from the group consisting of seq id no:

(a) (I) SEQ ID NO 52(HCDR1), SEQ ID NO 53(HCDR2), SEQ ID NO 54(HCDR3), SEQ ID NO 65(LCDR1), SEQ ID NO 66(LCDR2) and SEQ ID NO 67(LCDR 3); (II) SEQ ID NO:55(HCDR1), SEQ ID NO:53(HCDR2), SEQ ID NO:54(HCDR3), SEQ ID NO:65(LCDR1), SEQ ID NO:66(LCDR2) and SEQ ID NO:67(LCDR 3); (III) SEQ ID NO:56(HCDR1), SEQ ID NO:57(HCDR2), SEQ ID NO:54(HCDR3), SEQ ID NO:68(LCDR1), SEQ ID NO:69(LCDR2), and SEQ ID NO:70(LCDR 3); or (IV) SEQ ID NO 58(HCDR1), SEQ ID NO 59(HCDR2), SEQ ID NO 60(HCDR3), SEQ ID NO 71(LCDR1), SEQ ID NO 69(LCDR2) and SEQ ID NO 67(LCDR 3);

(b) (I) SEQ ID NO:76(HCDR1), SEQ ID NO:77(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:89(LCDR1), SEQ ID NO:90(LCDR2) and SEQ ID NO:91(LCDR 3); (II) SEQ ID NO:79(HCDR1), SEQ ID NO:77(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:89(LCDR1), SEQ ID NO:90(LCDR2) and SEQ ID NO:91(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:81(HCDR2), SEQ ID NO:78(HCDR3), SEQ ID NO:92(LCDR1), SEQ ID NO:93(LCDR2), and SEQ ID NO:94(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 83(HCDR2), SEQ ID NO 84(HCDR3), SEQ ID NO 95(LCDR1), SEQ ID NO 93(LCDR2) and SEQ ID NO 91(LCDR 3);

(c) (I) SEQ ID NO:310(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2) and SEQ ID NO:361(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:361(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:362(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 314(HCDR2), SEQ ID NO 349(HCDR3), SEQ ID NO 326(LCDR1), SEQ ID NO 324(LCDR2) and SEQ ID NO 361(LCDR 3);

(d) (I) SEQ ID NO:270(HCDR1), SEQ ID NO:389(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (II) SEQ ID NO:273(HCDR1), SEQ ID NO:389(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:390(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 275(HCDR1), SEQ ID NO 391(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3);

(e) (I) SEQ ID NO:407(HCDR1), SEQ ID NO:408(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (II) SEQ ID NO:409(HCDR1), SEQ ID NO:408(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:410(HCDR1), SEQ ID NO:411(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 412(HCDR1), SEQ ID NO 413(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3);

(f) (I) SEQ ID NO:310(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:321(LCDR2) and SEQ ID NO:322(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:321(LCDR2), and SEQ ID NO:322(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:312(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2), and SEQ ID NO:325(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 314(HCDR2), SEQ ID NO 315(HCDR3), SEQ ID NO 326(LCDR1), SEQ ID NO 324(LCDR2) and SEQ ID NO 322(LCDR 3);

(g) (I) SEQ ID NO 270(HCDR1), SEQ ID NO 271(HCDR2), SEQ ID NO 331(HCDR3), SEQ ID NO 337(LCDR1), SEQ ID NO 338(LCDR2) and SEQ ID NO 339(LCDR 3); (II) SEQ ID NO:273(HCDR1), SEQ ID NO:271(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:337(LCDR1), SEQ ID NO:338(LCDR2) and SEQ ID NO:339(LCDR 3); (III) SEQ ID NO:32(HCDR1), SEQ ID NO:274(HCDR2), SEQ ID NO:331(HCDR3), SEQ ID NO:340(LCDR1), SEQ ID NO:341(LCDR2), and SEQ ID NO:342(LCDR 3); or (IV) SEQ ID NO 275(HCDR1), SEQ ID NO 276(HCDR2), SEQ ID NO 332(HCDR3), SEQ ID NO 343(LCDR1), SEQ ID NO 341(LCDR2) and SEQ ID NO 339(LCDR 3); or

(h) (I) SEQ ID NO:310(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2) and SEQ ID NO:355(LCDR 3); (II) SEQ ID NO:229(HCDR1), SEQ ID NO:311(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:320(LCDR1), SEQ ID NO:354(LCDR2), and SEQ ID NO:355(LCDR 3); (III) SEQ ID NO:80(HCDR1), SEQ ID NO:313(HCDR2), SEQ ID NO:348(HCDR3), SEQ ID NO:323(LCDR1), SEQ ID NO:324(LCDR2) and SEQ ID NO:356(LCDR 3); or (IV) SEQ ID NO 82(HCDR1), SEQ ID NO 314(HCDR2), SEQ ID NO 349(HCDR3), SEQ ID NO 326(LCDR1), SEQ ID NO 324(LCDR2) and SEQ ID NO 355(LCDR 3).

Example 36 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of embodiments 1-5, 7-20, 26, 28, 30, 32, or 34, wherein the antibody or antigen-binding fragment comprises:

(a) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 136;

(b) 122, and a light chain variable region comprising the amino acid sequence of SEQ ID No. 136;

(c) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 128;

(d) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 128;

(e) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 147;

(f) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 147;

(g) (ii) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO:201 and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 174;

(h) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 174;

(i) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 180;

(j) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO. 180;

(k) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 201 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 186;

(l) 122, and a light chain variable region comprising the amino acid sequence of SEQ ID NO 186;

(m) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:103 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24;

(n) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:115 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24;

(o) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:122 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 48;

(p) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 128;

(q) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 136;

(r) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 147;

(s) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:154 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24;

(t) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:161 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24;

(u) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:168, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24;

(v) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 174;

(w) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 180;

(x) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO 37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 186;

(y) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:193 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 136;

(z) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:193, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 174;

(aa) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:13 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 24;

(bb) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:37, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 48;

(cc) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:374, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 244;

(dd) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:385 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 244;

(ee) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO:233, and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 244;

(ff) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO:278 and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 287; or

(gg) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:300 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 306.

Example 37. an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of

embodiments

1, 2, 4, 6, 21-25, 27, 29, 31, 33, or 35, wherein the antibody or antigen-binding fragment comprises:

(a) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO 61 and the light chain variable region comprising the amino acid sequence of SEQ ID NO 72;

(b) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 85 and a light chain variable region comprising the amino acid sequence of SEQ ID NO 96;

(c) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:350 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 363;

(d) 392 and 344;

(e) the heavy chain variable region comprising the amino acid sequence of SEQ ID NO. 414 and the light chain variable region comprising the amino acid sequence of SEQ ID NO. 344;

(f) 316, and a light chain variable region comprising the amino acid sequence of SEQ ID No. 327;

(g) (ii) a heavy chain variable region comprising the amino acid sequence of SEQ ID No. 333 and a light chain variable region comprising the amino acid sequence of SEQ ID No. 344; or

(h) A heavy chain variable region comprising the amino acid sequence of SEQ ID NO:350 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 357.

Example 38, an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of embodiments 1-5, 7-20, 26, 28, 30, 32, 34, or 36, wherein the antibody or antigen-binding fragment comprises:

(a) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 138;

(b) a heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 138;

(c) A heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 130;

(d) a heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 130;

(e) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 149;

(f) a heavy chain comprising the amino acid sequence of SEQ ID NO:208 and a light chain comprising the amino acid sequence of SEQ ID NO: 149;

(g) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 176;

(h) a heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 176;

(i) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 182;

(j) a heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 182;

(k) a heavy chain comprising the amino acid sequence of SEQ ID NO. 203 and a light chain comprising the amino acid sequence of SEQ ID NO. 188;

(l) A heavy chain comprising the amino acid sequence of SEQ ID NO 208 and a light chain comprising the amino acid sequence of SEQ ID NO 188;

(m) a heavy chain comprising the amino acid sequence of SEQ ID NO:105 and a light chain comprising the amino acid sequence of SEQ ID NO: 26;

(n) a heavy chain comprising the amino acid sequence of SEQ ID NO:108 and a light chain comprising the amino acid sequence of SEQ ID NO: 26;

(o) a heavy chain comprising the amino acid sequence of SEQ ID NO:117 and a light chain comprising the amino acid sequence of SEQ ID NO: 26;

(p) a heavy chain comprising the amino acid sequence of SEQ ID NO:124 and a light chain comprising the amino acid sequence of SEQ ID NO: 50;

(q) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 130;

(r) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 138;

(s) a heavy chain comprising the amino acid sequence of SEQ ID NO:141 and a light chain comprising the amino acid sequence of SEQ ID NO: 138;

(t) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 149;

(u) a heavy chain comprising the amino acid sequence of SEQ ID NO:156 and a light chain comprising the amino acid sequence of SEQ ID NO: 26;

(v) a heavy chain comprising the amino acid sequence of SEQ ID NO. 159 and a light chain comprising the amino acid sequence of SEQ ID NO. 26;

(w) a heavy chain comprising the amino acid sequence of SEQ ID NO:163 and a light chain comprising the amino acid sequence of SEQ ID NO: 26;

(x) A heavy chain comprising the amino acid sequence of SEQ ID NO:170 and a light chain comprising the amino acid sequence of SEQ ID NO: 26;

(y) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 176;

(z) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 182;

(aa) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 188;

(bb) a heavy chain comprising the amino acid sequence of SEQ ID NO:195, and a light chain comprising the amino acid sequence of SEQ ID NO: 138;

(cc) a heavy chain comprising the amino acid sequence of SEQ ID NO:195, and a light chain comprising the amino acid sequence of SEQ ID NO: 176;

(dd) a heavy chain comprising the amino acid sequence of SEQ ID NO:15 and a light chain comprising the amino acid sequence of SEQ ID NO: 26;

(ee) a heavy chain comprising the amino acid sequence of SEQ ID NO:39, and a light chain comprising the amino acid sequence of SEQ ID NO: 50;

(ff) a heavy chain comprising the amino acid sequence of SEQ ID NO:376 and a light chain comprising the amino acid sequence of SEQ ID NO: 246;

(gg) a heavy chain comprising the amino acid sequence of SEQ ID NO:387 and a light chain comprising the amino acid sequence of SEQ ID NO: 246;

(hh) a heavy chain comprising the amino acid sequence of SEQ ID NO:235 and a light chain comprising the amino acid sequence of SEQ ID NO: 246;

(ii) a heavy chain comprising the amino acid sequence of SEQ ID NO 280 and a light chain comprising the amino acid sequence of SEQ ID NO 289; or

(jj) a heavy chain comprising the amino acid sequence of SEQ ID NO:302, and a light chain comprising the amino acid sequence of SEQ ID NO: 308.

Example 39 an isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of

embodiments

1, 2, 4, 6, 21-25, 27, 29, 31, 33, 35, or 37, wherein the antibody or antigen-binding fragment comprises:

(a) a heavy chain comprising the amino acid sequence of SEQ ID NO 63 and a light chain comprising the amino acid sequence of SEQ ID NO 74;

(b) a heavy chain comprising the amino acid sequence of SEQ ID NO:87 and a light chain comprising the amino acid sequence of SEQ ID NO: 98;

(c) a heavy chain comprising the amino acid sequence of SEQ ID NO 352 and a light chain comprising the amino acid sequence of SEQ ID NO 365;

(d) a heavy chain comprising the amino acid sequence of SEQ ID NO. 394 and a light chain comprising the amino acid sequence of SEQ ID NO. 346;

(e) A heavy chain comprising the amino acid sequence of SEQ ID NO 416, and a light chain comprising the amino acid sequence of SEQ ID NO 346;

(f) a heavy chain comprising the amino acid sequence of SEQ ID NO:318, and a light chain comprising the amino acid sequence of SEQ ID NO: 329;

(g) a heavy chain comprising the amino acid sequence of SEQ ID NO 335 and a light chain comprising the amino acid sequence of SEQ ID NO 346; or

(h) A heavy chain comprising the amino acid sequence of SEQ ID NO 352 and a light chain comprising the amino acid sequence of SEQ ID NO 359.

Example 40. the antibody or antigen-binding fragment of any one of examples 1-39, which is an antigen-binding fragment selected from the group consisting of: fab, Fab ', F (ab')₂Fv, single domain antibody (dAb), and single chain variable fragment (scFv), optionally wherein the antigen binding fragment is selected from the group consisting of: fab, Fab', Fv, single domain antibody (dAb), and single chain variable fragment (scFv).

Example 41. the antibody or antigen-binding fragment of any one of examples 1-40, which is monoclonal.

Embodiment 42. the antibody or antigen-binding fragment of any one of embodiments 1-41, which is fully human.

Embodiment 43 the antibody or antigen binding fragment of any one of embodiments 1-42, which is an IgG antibody, optionally an IgG1 antibody.

Example 44. the antibody or antigen-binding fragment of any one of examples 1-43, which is an IgG1 antibody having a kappa light chain.

Example 45 the antibody or antigen binding fragment of any one of examples 1-44, which is a fully human antibody of the IgG1 isotype and has a kappa light chain.

Embodiment 46. the antibody or antigen-binding fragment of any one of embodiments 1-45, wherein the antibody or antigen-binding fragment has further modifications as described herein, e.g., wherein the antibody or antigen-binding fragment additionally has a mutation in the Fc region according to the EU index of kabat, wherein the mutation comprises at least D265A and P329A; or wherein the mutation comprises at least L234A and L235A.

Embodiment 47. the antibody or antigen-binding fragment of any one of embodiments 1-40, wherein the antibody or antigen-binding fragment is: a) is monoclonal; and/or b) fully human; and/or c) an IgG antibody, optionally an IgG1 antibody; and/or d) having a kappa light chain; and/or e) has a mutation in the Fc region according to EU index of kabat, optionally wherein the mutation comprises at least D265A and P329A; and/or f) has a mutation in the Fc region according to the EU index of kabat, optionally wherein the mutation comprises at least L234A and L235A.

Embodiment 48 the antibody or antigen-binding fragment of any one of embodiments 1-47, wherein the antibody or antigen-binding fragment is therapeutic.

Example 49. an isolated antibody or antigen-binding fragment that binds to the same epitope on human NPR1 as the antibody or antigen-binding fragment of any one of examples 1 to 48.

Example 50 an isolated antibody or antigen-binding fragment that competes for binding to human NPR1 with the antibody or antigen-binding fragment of any one of examples 1 to 49.

Example 51, one or more isolated nucleic acids encoding the amino acid sequence of an antibody or antigen-binding fragment of any one of examples 1 to 50.

Example 52A vector comprising one or more isolated nucleic acids as described in example 51.

Example 53. a host cell comprising one or more isolated nucleic acids as described in example 51 or a vector as described in example 52.

Example 54. a method of producing the antibody or antigen-binding fragment of any one of examples 1 to 50, comprising culturing the host cell of example 53 under conditions suitable for production of the antibody or antigen-binding fragment.

Embodiment 55. the method of embodiment 54, wherein the method additionally comprises purifying the antibody or antigen binding fragment.

Example 56 a pharmaceutical composition comprising a purified antibody or antigen-binding fragment produced by the method of example 55, and a pharmaceutically acceptable carrier.

Embodiment 57 a pharmaceutical composition comprising the antibody or antigen-binding fragment of any one of embodiments 1 to 50, and a pharmaceutically acceptable carrier.

Embodiment 58. the pharmaceutical composition of embodiment 56 or 57 or the combination comprising the antibody or antigen-binding fragment of any one of embodiments 1 to 50, wherein the composition further comprises an additional therapeutic agent.

Embodiment 59. the pharmaceutical composition or combination of embodiment 58, wherein the additional therapeutic agent is selected from ACE (angiotensin converting enzyme) inhibitors, Angiotensin Receptor Blockers (ARBs), enkephalinase inhibitors, beta blockers, diuretics, calcium channel blockers, cardiac glycosides, sodium-glucose cotransporter 2 inhibitors (SGLT2i), and combinations thereof.

Example 60. the pharmaceutical composition or combination of example 58 or example 59, wherein the additional therapeutic agent is selected from enalapril, benazepril, captopril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, trandolapril, valsartan, azilsartan, candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, sacubitril, bisoprolol, carvedilol, propranolol, metoprolol tartrate, metoprolol succinate, thiazide diuretics, ring diuretics, potassium sparing diuretics, amlodipine, clevidipine, diltiazem, felodipine, isradipine, nicardipine, nifedipine, nisoldipine, verapamil, diguanoside, canagliflozin, dacliptin, eprolean, epilagliptin, anglezin, anglegliflozin, and mixtures thereof, And combinations thereof.

Embodiment 61. the pharmaceutical composition or combination of embodiment 58 or embodiment 59, wherein the additional therapeutic agent is selected from the group consisting of chlorothiazide, chlorthalidone, hydrochlorothiazide, indapamide, metolazone, bumetanide, ethacrynic acid, furosemide, torasemide, amiloride, eplerenone, spironolactone, triamterene, digoxin, and combinations thereof.

Embodiment 62. the pharmaceutical composition or combination of any one of embodiments 58-61, wherein the additional therapeutic agent is an angiotensin receptor-enkephalinase inhibitor (ARNi).

Embodiment 63. the pharmaceutical composition or combination of embodiment 58, wherein the additional therapeutic agent is selected from the group consisting of a corticosteroid, a leukotriene modifier, a bronchodilator, and combinations thereof.

Embodiment 64. the pharmaceutical composition or combination of embodiment 63, wherein the additional therapeutic agent is selected from the group consisting of fluticasone, budesonide, mometasone, beclomethasone, ciclesonide, fluticasone furoate, prednisone, methylprednisolone, montelukast, zafirlukast, zileuton, long-acting beta agonists, short-acting beta agonists, theophylline and ipratropium, and combinations thereof.

Embodiment 65. the pharmaceutical composition or combination of embodiment 63 or embodiment 64, wherein the additional therapeutic agent is selected from the group consisting of salmeterol, formoterol, salbutamol and levalbuterol, and combinations thereof.

Embodiment 66. the pharmaceutical composition or combination of embodiment 58, wherein the additional therapeutic agent is selected from the group consisting of a β -adrenoreceptor antagonist, a carbonic anhydrase inhibitor, an α 2-adrenoreceptor agonist, a parasympathomimetic, a prostaglandin analog, a rho kinase inhibitor, and combinations thereof.

Embodiment 67. the pharmaceutical composition or combination of embodiment 66, wherein the additional therapeutic agent is selected from the group consisting of timolol, levobunolol, metiprolol, carteolol, betaxolol, acetazolamide, dorzolamide, brinzolamide, methazolamide, brimonidine, alaclonidine, a cholinomimetic, latanoprost, latanoproste bund, travoprost, bimatoprost, tafluprost, neratesudil, and lapachladide, and combinations thereof.

Embodiment 68 the pharmaceutical composition of embodiment 58, wherein the additional therapeutic agent is selected from ACE (angiotensin converting enzyme) inhibitors, Angiotensin Receptor Blockers (ARBs), enkephalinase inhibitors, beta blockers, diuretics, calcium channel blockers, cardiac glycosides, sodium-glucose cotransporter 2 inhibitors (SGLT2i), angiotensin receptor-enkephalinase inhibitors (ARNi), corticosteroids, leukotriene modifiers, bronchodilators, beta-adrenoceptor antagonists, carbonic anhydrase inhibitors, alpha 2-adrenoceptor agonists, parasympathomimetics, prostaglandin analogs, rho kinase inhibitors, and combinations thereof.

Example 69. the pharmaceutical composition of example 68, wherein the additional therapeutic agent is selected from enalapril, benazepril, captopril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, trandolapril, valsartan, azilsartan, candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, sabotatrazole, bisoprolol, carvedilol, propranolol, metoprolol tartrate, metoprolol succinate, thiazide diuretics, periantidiuretic, amlodipine, clevidipine, diltiazem, felodipine, isradipine, nicardipine, nifedipine, nisoldipine, verapamil, digitoxin, canagliflozin, dacagliflozin, eprogliflozin, ethiazide, chlorothiazide, losterone, lostazidone, ramipril, and ramipril, Hydrochlorothiazide, indapamide, metolazone, bumetanide, ethacrynic acid, furosemide, torasemide, amiloride, eplerenone, spironolactone, triamterene, digoxin, fluticasone, budesonide, mometasone, beclomethasone, ciclesonide, fluticasone furoate, prednisone, methylprednisolone, montelukast, zafirlukast, zileuton, long-acting beta agonists, short-acting beta agonists, theophylline, ipratropium, salmeterol, formoterol, salbutamol, and levalbuterol, timolol, levobunolol, metiprolol, betaxolol, acetazolamide, dorzolamide, brinzolamide, methazolamide, brimonidine, aclonidine, prostaglandins, latanoprost (latanoprost), latanoprost, breglist, brevoprost, brevone, brevoxamine, flunomide, doxepirubigin, doxepirubine, doxepirubigin, valbutirolatanolide, doxylamine, latanolide, doxine, latanolide, latanoprost, latanolide, flunomide, flunomil, fludarabine, fludarmerely, fludarabine, trolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolatanolat, Neratesudil and lapacholide, and combinations thereof.

Example 70 the antibody or antigen-binding fragment thereof of any one of examples 1-50, the one or more isolated nucleic acids of example 51, the vector of example 52, the host cell of example 53, or the pharmaceutical composition of any one of examples 56-69, for use (i) in therapy, (ii) as a medicament, or (iii) in the manufacture of a medicament for treating a disease.

Example 71 use of the antibody or antigen binding fragment of any one of examples 1-50, one or more isolated nucleic acids of example 51, a vector of example 52, a host cell of example 53, or a pharmaceutical composition of any one of examples 56-69, for the manufacture of a medicament for treating a disorder or disease associated with natriuretic peptide receptor activity in a subject in need of such treatment.

Example 72 use of the antibody or antigen-binding fragment of any one of examples 1-50, one or more isolated nucleic acids of example 51, a vector of example 52, a host cell of example 53, or a pharmaceutical composition of any one of examples 56-69, for the manufacture of a medicament for treating a cardiovascular disorder in a subject in need of such treatment.

Embodiment 73 the use of embodiment 72, wherein the cardiovascular disorder is selected from the group consisting of: hypertension, peripheral vascular disease, heart failure, Coronary Artery Disease (CAD), Ischemic Heart Disease (IHD), mitral stenosis and regurgitation, angina, hypertrophic cardiomyopathy, diabetic cardiomyopathy, supraventricular and ventricular arrhythmias, arrhythmia, Atrial Fibrillation (AF), new atrial fibrillation, recurrent atrial fibrillation, cardiac fibrosis, atrial flutter, detrimental vascular remodeling, plaque stabilization and Myocardial Infarction (MI).

Example 74 use of the antibody or antigen-binding fragment of any one of examples 1-50, one or more isolated nucleic acids of example 51, a vector of example 52, a host cell of example 53, or a pharmaceutical composition of any one of examples 56-69, for the manufacture of a medicament for treating heart failure, Hypertrophic Cardiomyopathy (HCM), hypertension, pre-eclampsia, asthma, glaucoma, and/or cytokine release syndrome in a subject in need of such treatment.

Example 75 the use of example 73 or example 74, wherein the subject has heart failure, and wherein the heart failure is selected from heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), post-acute myocardial infarction heart failure, or acute compensated heart failure.

Embodiment 76 the use of embodiment 73 or embodiment 74, wherein the subject has hypertrophic cardiomyopathy, and wherein the hypertrophic cardiomyopathy is ventricular hypertrophy.

Embodiment 77 the use of embodiment 73 or embodiment 74, wherein the subject has hypertension, and wherein the hypertension is selected from refractory hypertension, hypertensive heart disease, pulmonary hypertension, pulmonary arterial hypertension, isolated systolic hypertension, refractory hypertension, and pulmonary arterial hypertension.

Embodiment 78 the use of embodiment 73, 74, or 77, wherein the subject has hypertension, and wherein the hypertension is selected from refractory hypertension or hypertensive heart disease.

Example 79 use of the antibody or antigen binding fragment of any one of examples 1-50, one or more isolated nucleic acids of example 51, a vector of example 52, a host cell of example 53, or a pharmaceutical composition of any one of examples 56-69, for the manufacture of a medicament for treating a renal disorder in a subject in need of such treatment.

Embodiment 80 the use of embodiment 79, wherein the renal disorder is selected from the group consisting of: diabetic renal insufficiency, non-diabetic renal insufficiency, renal failure, diabetic nephropathy, non-diabetic nephropathy, acute kidney injury, contrast-induced nephropathy, nephrotic syndrome, glomerulonephritis, scleroderma, glomerulosclerosis, primary nephrotic proteinuria, renovascular hypertension, diabetic retinopathy and end-stage renal disease (ESRD), endothelial dysfunction, diastolic dysfunction, renal fibrosis and Polycystic Kidney Disease (PKD).

Example 81 the antibody or antigen-binding fragment of any one of examples 1-50, the one or more isolated nucleic acids of example 51, the vector of example 52, the host cell of example 53, or the pharmaceutical composition or combination of any one of examples 56-69 for use in treating a disorder or disease associated with natriuretic peptide receptor activity in a subject in need of such treatment.

Example 82 the antibody or antigen-binding fragment of any one of examples 1-50, the one or more isolated nucleic acids of example 51, the vector of example 52, the host cell of example 53, or the pharmaceutical composition or combination of any one of examples 56-69, are used to treat a cardiovascular disorder in a subject in need of such treatment.

Example 83 the antibody or antigen binding fragment, one or more isolated nucleic acids, vector, host cell, pharmaceutical composition, or combination of example 82, wherein the cardiovascular disorder is selected from the group consisting of: hypertension, peripheral vascular disease, heart failure, Coronary Artery Disease (CAD), Ischemic Heart Disease (IHD), mitral stenosis and regurgitation, angina, hypertrophic cardiomyopathy, diabetic cardiomyopathy, supraventricular and ventricular arrhythmias, arrhythmia, Atrial Fibrillation (AF), new atrial fibrillation, recurrent atrial fibrillation, cardiac fibrosis, atrial flutter, detrimental vascular remodeling, plaque stabilization and Myocardial Infarction (MI).

Example 84 the antibody or antigen-binding fragment of any one of examples 1-50, the one or more isolated nucleic acids of example 51, the vector of example 52, the host cell of example 53, or the pharmaceutical composition or combination of any one of examples 56-69, for use in treating heart failure, Hypertrophic Cardiomyopathy (HCM), hypertension, preeclampsia, asthma, glaucoma, and/or cytokine release syndrome in a subject in need of such treatment.

Example 85 the antibody or antigen-binding fragment, one or more isolated nucleic acids, vector, host cell, pharmaceutical composition, or combination of example 83 or example 84, wherein the subject has heart failure, and wherein the heart failure is selected from the group consisting of heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), heart failure after acute myocardial infarction, or acute compensated heart failure.

Example 86 the antibody or antigen-binding fragment, one or more isolated nucleic acids, vector, host cell, pharmaceutical composition, or combination of example 83 or example 84, wherein the subject has hypertrophic cardiomyopathy, and wherein the hypertrophic cardiomyopathy is ventricular hypertrophy.

The antibody or antigen-binding fragment, one or more isolated nucleic acids, vector, host cell, pharmaceutical composition, or combination of example 83 or example 84, wherein the subject has hypertension, and wherein the hypertension is selected from refractory hypertension, hypertensive heart disease, pulmonary hypertension, pulmonary arterial hypertension, isolated systolic hypertension, refractory hypertension, and pulmonary arterial hypertension.

Example 88 the antibody or antigen binding fragment, one or more isolated nucleic acids, vector, host cell, pharmaceutical composition, or combination of examples 83, 84, or 87, wherein the subject has hypertension, and wherein the hypertension is selected from refractory hypertension or hypertensive heart disease.

Example 89 the antibody or antigen-binding fragment of any one of examples 1-50, the one or more isolated nucleic acids of example 51, the vector of example 52, the host cell of example 53, or the pharmaceutical composition or combination of any one of examples 56-69, is used to treat a renal disorder in a subject in need of such treatment.

Example 90 the antibody or antigen binding fragment, one or more isolated nucleic acids, vector, host cell, pharmaceutical composition, or combination of example 89, wherein the renal disorder is selected from the group consisting of: diabetic renal insufficiency, non-diabetic renal insufficiency, renal failure, diabetic nephropathy, non-diabetic nephropathy, acute kidney injury, contrast-induced nephropathy, nephrotic syndrome, glomerulonephritis, scleroderma, glomerulosclerosis, primary nephrotic proteinuria, renovascular hypertension, diabetic retinopathy and end-stage renal disease (ESRD), endothelial dysfunction, diastolic dysfunction, renal fibrosis and Polycystic Kidney Disease (PKD).

Examples of the invention

The following examples provide illustrative embodiments of the present disclosure. Those of ordinary skill in the art will recognize that various modifications and changes can be made without changing the spirit or scope of the present disclosure. Such modifications and variations are intended to be included within the scope of the present disclosure. The examples provided do not limit the disclosure in any way.

The disclosure provides anti-NPR 1 antibodies that specifically bind to and activate NPR1, e.g., (i) bind to NPR 1; and (ii) activates an antibody or antigen-binding fragment of NPR1 in the absence of ANP. Antibodies that specifically bind to and activate NPR1 may have different possible modes of action: (1) the antibody induces a conformational change in the NPR1 monomer to activate the receptor; (2) the antibody directly mimics the structure and function of the natural ligand ANP and activates the receptor by binding in the ANP binding pocket of NPR 1; or (3) the antibody stabilizes a preformed functionally active complex of hNPR1 and ANP (NPR1-ANP complex).

Example 1:

phage library panning and screening

To select NPR 1-specific antibodies encompassing the different modes of action described, 13 different panning strategies were applied (see table 5). Only ten strategies were performed on the protein (strategies 1-6 and 10-13). In addition, three different cell elutriations were performed (strategies 7-9). A total of four panning strategies (strategies 3 and 11-13) were aimed at enriching for ANP competitive antibodies (elution with ANP, pre-adsorption of phage on NPR1-ANP complex, and anti-idiotypic panning on murine anti-ANP antibodies).

Table 5.

Overview of panning strategy

For Fc capture panning, NPR1-hFc was immobilized on 96-well plates by an appropriate capture antibody (goat or mouse anti-human Fc antibody). The antigen was fixed in the appropriate number of wells of a 96-well plate, and the wells were then blocked prior to the addition of phage antibody. In parallel to well preparation, phage antibodies were blocked. During blocking of the phage, additional blocking reagents were added to the blocking buffer to avoid selection of antibodies against the hFc tag or capture antibody (goat or mouse gamma globulin). Following the blocking procedure, two pre-adsorption steps were performed on human gamma globulin and the reverse target hNPR3-hFc to avoid selection of antibodies against the Fc-tag or the reverse target. The pre-blocked and pre-adsorbed phage mixture with immobilized NPR1-hFc was added to each well and phage-antibodies were allowed to bind to the antigen. The intensive washing ensures removal of non-specifically bound phage and then elution of specifically bound phage. The second and third rounds of solid phase panning were performed according to the protocol of the first round of panning. The amount of antigen is reduced and washing conditions with increased stringency are applied.

For solution panning, NPR1 was biotinylated and the retained activity of biotinylated NPR1 on ANP binding was confirmed. During solution panning, Fab-displaying phage and biotinylated NPR1-hFc were incubated in solution, which facilitated phage access to the antigen. Blocking an appropriate amount of streptavidin beads and simultaneously blocking an appropriate amount of phage antibodies. During phage blocking, human gamma globulin, the reverse target hNPR3-hFc and Flag-TEV linker peptide were added to the blocking buffer to avoid selection of antibodies against the hFc tag, the reverse target or the linker peptide. To remove streptavidin, biotin or bead-bound phage, a pre-adsorption step of blocked phage particles is performed using blocked streptavidin beads with or without conjugated biotinylated unrelated antigens. Thus, biotinylated NPR1-hFc/NPR1-hFc-ANP complex was added to the pre-adsorbed and blocked phage particles and allowed the phage-antibody to bind antigen in solution. To enrich for antibody phages binding to the ANP binding site of NPR1 (ANP competitive antibodies), a preformed NPR1-ANP complex was added to the phage blocking solution, or ANP peptide was used to elute bound phages. Thus, at least a 250-fold molar excess of the ANP peptide used over the NPR1 antigen or NPR1 expressing cells was used. The blocked streptavidin beads were used to capture phage-antigen complexes, and the streptavidin bead-bound phage particles were collected with a magnetic separator. Non-specifically bound phage are washed away by several washing steps. Specifically bound phage are eluted from the streptavidin beads. The eluate was transferred to E.coli cultures for phage infection. The second and third rounds of bead-based solution panning were performed according to the protocol of the first round panning. The amount of antigen is reduced and washing conditions with increased stringency are applied.

For whole cell panning, appropriate amounts of phage-antibody were blocked. During phage blocking, the reverse target hNPR3-hFc was added to the blocking buffer to avoid selecting antibodies against the reverse target. At the same time, each phage pool blocked an appropriate number of target cells expressing NPR1 and an appropriate number of adsorbed cells not expressing antigen (parental cells). Blocked target cells were spun down, resuspended in pre-blocked phage particles, and phage-antibodies were allowed to bind to NPR1 presented on the cells. The phage-cell complex was washed several times. To enrich for antibody phages binding to the ANP binding site of NPR1 (ANP competitive antibodies), a preformed NPR1-ANP complex was added to the phage blocking solution, or ANP peptide was used to elute bound phages. Thus, at least a 250-fold molar excess of the ANP peptide used over the NPR1 antigen or NPR1 expressing cells was used. Specifically bound phage are eluted from the target cells. After centrifugation, the supernatant (eluate) is applied to the adsorbed cells to remove phage bound to cell surface molecules other than the target antigen (after adsorption). The final supernatant was transferred to E.coli cultures for phage infection. The second and third rounds of whole cell panning were performed according to the protocol of the first round of panning. Washing conditions with increased stringency are applied.

The output of the panning wheel was then subcloned into bacterial expression vectors and bacterial lysates (BEL) were used for primary and secondary screens. During primary screening (ELISA-based), the output was analyzed for binding to human and rat NPR 1. Clones that bound to human NPR3 were deleted. Secondary screening was performed on CHO-K1 cells expressing hNPR 1. Further screening for ANP competition and binding was performed in the presence of ANP only. Approximately 1700 clones met the selection criteria for screening and 760 were selected for sequencing. Sequencing of 760 clones yielded 210 unique hits of HCDR3, the binding characteristics of which are summarized in table 6. Of these clones, 72 showed significant ANP competition, while 7 clones only bound in the presence of ANP.

TABLE 6.210 HCDR3 uniquenessHit in

In the binding characteristics of

Example 2: antibody reformatting, expression and purification

After confirmation of binding, 210 unique cloned VH and VL domains of HCDR3 were subcloned into a vector with human IgG constant regions. 180 out of 210 clones were selected for expression, and 166 out of 180 passed production quality control. They were characterized in terms of binding to relevant cell lines and functional activity. Then 40 out of 166 candidates were selected for exploratory scale production, 31 of these candidates were characterized in detail in terms of binding to relevant antigens and cell lines, ANP competition and functionality in a cell-based cGMP production assay, as shown below.

To generate IgG candidates, eukaryotic HKB11 cells were transfected with mammalian expression vector DNA encoding the heavy and light chains of IgG. Cell culture supernatants were harvested at appropriate times and subjected to protein a affinity chromatography. If necessary, a second purification step is performed to remove aggregates. The buffer was exchanged into 1 × Duchen PBS (pH 7.2) and the sample was sterile filtered (0.2 μm pore size).

Example 3: antigen binding, ANP competition and cellular cGMP production-

Candidates

31 IgG, quality controlled by exploratory scale production, were tested for binding to the following antigens via ELISA: human NPR1, constitutively active human NPR1 mutant (W74R), rat NPR1 and human NPR3 (reverse target). Clones were also tested by flow cytometry for binding to CHO K1 cells expressing human NPR1 in the absence and presence of ANP, as well as on parental CHO K1 cells. The binding properties of the five functional candidates are shown in fig. 1 and 2.

The same 31 IgG were tested for ANP competition using a Fluorescence Resonance Energy Transfer (FRET) based assay, in which NPR 1-specific antibodies competed with ANP for binding to NPR 1. In this FRET-based assay (see fig. 3), Eu-labeled streptavidin (for measurement of IgG) or Eu-labeled anti-hFc antibody (for measurement of FabCys) was used as energy donor, while Cy 5-labeled ANP was used as acceptor (for all measurements). The generated fluorescent signal is reduced by ANP competitive antibodies. The assay was performed as follows: the antibodies were mixed with NPR1 and incubated for 5 minutes at room temperature. After adding Eu-labeled donor and incubation at room temperature for 30 min, Cy5-ANP solution was added. After an additional 60 minutes of incubation, readings were taken using TECAN Infinite M1000 Pro (using an excitation wavelength of 317nm and an emission wavelength of 665 nm). The percentage of ANP competition was calculated according to the following formula:

Ratio [ (A)₆₆₅nm/A₆₂₀nm)*10⁴]

Ratio (ratio-ratio)_neg)

Competition% [100- (ratio/(ratio))_pos/100))]

Ratio of_neg: mean ratio data values for control without NPR1

Ratio of_pos: mean ratio data for control without agonist (reaction buffer)

15 of the 31 iggs were ANP competitive, but only two of these candidates showed functionality in the cGMP assay (WW04 and WW 06). The other three functional candidates WW01, WW02 and WW03 showed "negative" ANP competition in this assay, indicating stabilization of the NPR1-ANP complex. The results of the FRET assays for the five functional candidates are shown in FIG. 4.

In addition, functional activity of 31 IgG was tested in a cellular cGMP production assay using CHO-K1 cells expressing human NPR1, as described above. For functional characterization of the selected antibodies, the production of cyclic guanosine 3',5' -cyclic monophosphate (cGMP) was monitored after binding to and stimulation of NPR1 expressed on the cell surface of CHO-K1 cells. Cellular cGMP is a major second messenger that mediates cellular activity and is mediated by ANPOr NPR 1-specific antibody-triggered activated NPR1 synthesis. Therefore, a commercial assay kit (Cisbio Bioassays Cisbio HTRF assay kit Cisbio (catalog No. 62GM2PEB)) was used. The measurements were made according to the manufacturer's instructions (slight deviations). Briefly, cells were adjusted to 1X 10 ⁵cells/mL, 20 μ L/well were seeded in 96 well microtiter plates and incubated overnight. After addition of 10. mu.L/well of different concentrations of antibody, the plates were incubated at 37 ℃ for 30min to generate cGMP. In parallel, a standard curve was generated using a calibrator (contained in the kit). Cells were lysed and a mixture of cGMP-d2 and anti-cGMP-Cryptate (Cryptate) was added and incubated at room temperature for 1 h. Readings were taken using a Tecan M1000 Pro (using an excitation wavelength of 317nm and an emission wavelength of 665 nm). The cGMP concentration (Δ F [% ]) was calculated according to the following formula])：

Ratio [ (A)_665nm/B_620nm)*10⁴]

Average ratio ═ (Σ ratio/2)

CV [ ("standard deviation/mean value). 100]

Δ F [ ((calibrator or sample ratio-ratio)_neg) Ratio of_neg)*100]

Ratio of_neg: negative control

Five candidates with significant functional activity were identified using the cellular cGMP assay: WW01, WW02, WW03, WW04 and WW 06. These five candidates are functionally active and can be assigned to different modes of action. WW01, WW02 and WW03 were able to stabilize NPR1-ANP complex, whereas WW06 was determined to be ANP competitive. These candidates were all from initial panning codes 10 and 11 (for mode of action 2 or 3). Figure 5 shows the results of a cellular production assay of cGMP in the absence or presence of 0.075nM ANP induced by five functional candidates (IgG format).

Example 4: production and characterization of the FabCys form

Functional clones were also tested for functionality in the FabCys format. Eukaryotic HKB11 cells were transfected with mammalian expression vector DNA encoding the heavy and light chains of disulfide-bridged FabCys. Cell culture supernatant was collected at appropriate times and subjected to metal ion affinity chromatography using a liquid handling workstation. The buffer was exchanged into 1 × Duchen PBS (pH 7.2) and the sample was sterile filtered (0.2 μm pore size).

Five functional candidates WW01, WW02, WW03, WW04 and WW06 were additionally analyzed for their monovalent affinities for human and rat NPR1 and the reverse target human NPR3 in the absence and presence of ANP in the monovalent FabCys form. Table 7 summarizes the results of the affinity assay, epitope ranking and cGMP assay.

TABLE 7 summary of affinities, epitopes and cGMP data for WW01, WW02, WW03, WW04 and WW06 in the form of FabCys

For candidates WW01 and WW03, no or very weak binding to human and rat NPR1 was observed in the absence of ANP, whereas the affinity was in the low nanomolar to sub-nanomolar range in the presence of ANP. Both share the same epitope box "B". The affinity of the candidate WW02 was too weak to determine K adequately _DValues and epitope bins. The affinities of WW04 and WW06 ranged from two-digit nanomolar to subnanomolar, independent of the presence or absence of ANP. Both share the same epitope box "a". WW06 was the only candidate that did not exhibit rat cross-reactivity.

Although no binding to the reverse target hNPR3 was observed in the absence or presence of ANP for WW02, WW03, and WW04, additional binding to the reverse target was detected for both WW01 and higher concentrations of WW 06.

Example 5: will be provided with

Reformatted into IgG format

The FabCys vector was subcloned into the IgG1_ LALA vector for expression in mammalian cells by amplifying the Fab coding insert using one biotinylated primer and one non-biotinylated primer. The amplified product was bound to streptavidin beads, digested with restriction enzymes, and washed, releasing the purified insert into the supernatant. Inserts were cloned into recipient vectors, DNA was transformed, and individual clones were quality controlled by colony PCR and sequencing.

Five functional candidates in the IgG format, WW01, WW02, WW03, WW04 and WW06, were characterized as described above. Binding data (ELISA, flow cytometer, ANP competition) and functional data (cGMP assay) as well as affinity and epitope bins are shown in table 8. Interestingly, WW06 had significantly enhanced functional activity in the FabCys format compared to the IgG format, as shown in figure 6.

Table 8 summary of affinity, epitope and cGMP data for WW01, WW02, WW03, WW04 and WW06 in IgG format

Example 6:

generation of mature libraries

To increase the affinity and bioactivity of selected antibody fragments (WW01, WW02, WW03, WW04 and WW06), LCDR3 and HCDR2 regions were exchanged In parallel by diversified cassettes/modules (Prassler et al (2009): In vitro affinity mapping of

antibodies:complementarity determining region exchange and RapMAT technology[

In vitro affinity maturation of antibodies: CDRs exchange and RapMAT technology](ii) a Immunotherapy]1(4) page 571-583, which is described in detailThe contents of which are hereby incorporated by reference) while the framework regions remain constant. Parent Fab fragments were transferred from the corresponding expression vectors into library cloning vectors for affinity maturation.

Performed separately for each maturation candidate

Generation of a mature library. For LCDR3 optimization, an approximately 400bp DNA fragment encoding LCDR3, framework 4, and light chain constant region was removed from the parent antibody-encoding sequence by restriction digestion. To reduce the background of parental non-diversified sequences, the excised fragment was replaced by a virtual sequence of approximately 520bp by ligation, followed by insertion of a pool of DNA fragments encoding the diversified LCDR3 region with framework 4 and constant domains (diversified LCDR3 cassette) by restriction digestion and ligation.

In the second library set, the HCDR2 coding sequence was diversified, while the linker framework regions remained constant. To reduce the background of parental non-diversified sequences, an approximately 150bp DNA fragment containing the parental HCDR2 and framework 3 sequences was replaced with an approximately 590bp virtual sequence by restriction digestion and ligation, and then a diversified HCDR2 cassette (including framework 3) was also inserted by restriction digestion and ligation.

Ten mature libraries were successfully cloned and the library size was 9.2x10⁸And 2.2x10⁹Between cfu. Electroporation of the ligation mixture into E.coli cells results>10⁸Individual colonies. Amplification of the library was performed as described previously (Rauchenberger et al (2003): Human combinatorial Fab library derived specific and functional antibodies obtained the Human fibroblast growth factor receptor 3[ Human combinatorial Fab library generates specific and functional antibodies against Human fibroblast growth factor receptor 3](ii) a J Biol Chem [ journal of Biochemistry]278(40), pages 38194 and 38205, the contents of which are hereby incorporated by reference. For quality control, approximately 10-20 individual colonies per library were randomly picked and sequenced.

To select for improved affinity candidates, phages derived from the mature library were subjected to three rounds of maturation panning, as further described below. Extended washing steps can increase panning stringency. In addition, dissociation rate (off-rate) Selection was performed (Hawkins et al (1992): Selection of phase antibodies by binding affinity [ phage antibodies selected by binding affinity ] Mimicking affinity mapping [ mimic affinity maturation ] J.mol.biol. [ journal of molecular biology ]226 (3); page 889-.

Example 7: panning and screening-

The mature library was used for four different mature panning strategies. Strategies #3 and #4 are intended to enrich progeny with improved affinity compared to parental clones. In addition, strategies #1 and #2 were aimed at enriching clones with improved affinity for NPR1 but not for the NPR1-ANP complex. The rationale behind this is the idea of producing candidates that can directly activate NPR1 through conformational changes. During panning, all mature libraries were kept separately. The panning strategy is summarized in detail in table 9. The output of the third round of panning was then subcloned into bacterial expression vectors and bacterial lysates (BEL) used for SET screening.

Table 9:

overview of mature panning strategy

The output of the third round of panning was used for Solution Equilibrium Titration (SET) screening. The improved affinity of 88 clones/subcode (2640 clones in total) for hNPR1 and/or hNPR1-ANP complex was analyzed in the SET screen compared to the parental clones.

During SET screening, 82 HCDR2 or LCDR3 were identified as the only improved derivatives. Compared with their parent clones, the affinity of WW01 and WW03 derivatives to hNPR1 and hNPR1-ANP complex is improved by 20 times. The affinity of the WW04 derivative was not significantly improved compared to the parental clone, whereas the affinity of the WW06 derivative was improved by 3-fold. See FIG. 7 (shown as having affinity for hNPR 1) and FIG. 8 (shown as having affinity for hNPR1-ANP complex). Affinity (K) _D[pM]) Shown on the x-axis below the parent clone names of both figures.

Example 8: maturation candidates-

Is characterized by

Of the 82 improved candidates, 74 were successfully subcloned in the FabCys form, and 61 of the 74 clones passed production quality control and were characterized for binding to the relevant antigen, binding to the relevant cell line, ANP competition, and functional activity in cGMP production (compared to their parental clones). All 16 derivatives of WW01 and all 27 derivatives of WW03 had up to 20-fold improved binding and functional activity. Most derivatives also show improved binding and functionality in the presence of ANP. Some derivatives showed binding to W74R (constitutively active hNPR1 mutant), which was not the case for the parent FabCys. One of the four derivatives of WW04 had two-fold enhanced binding and functional activity, while the rest behaved like the parent FabCys. All 14 derivatives of WW06 improved binding to NPR1 and could remain competitive with ANP. Functional activity was increased three-fold in 10 of the 14 progeny compared to the parental FabCys. Some derivatives showed rat cross-reactivity, while the parental FabCys did not.

After FabCys characterization, another 40 derivatives were selected for IgG transformation and further characterization. Ten potential candidates shown in table 10 were then assayed for binding to the relevant antigen, binding to the relevant cell line, ANP competition, and functional activity in cGMP production (compared to their parental clones). They were further analyzed via 3P assay and via SET K_DMeasurements to determine their affinity to human and rat NPR1 in the absence and presence of ANP. WW01 and WW03 derivative antibodies were analyzed in IgG format and WW06 derivatives were analyzed in FabCys format.

For proteomic profiling (Freese et al (2013): An automated immunological profiling for early specificity analysis of antibodies; mAbs 5(2), page 279-287, the contents of which are hereby incorporated by reference, 32 different proteins and controls were coated at a concentration of 1.0. mu.g/mL overnight at 4 ℃ onto two 384-well MSD standard plates. The coating solution was discarded, the plate blocked with 50. mu.L of 3% (w/v) BSA in PBS on a microtiter plate shaker (about 500rpm) for 1 hour at room temperature, and then three wash steps were performed with 50. mu.L of wash buffer (PBS containing 0.05% (v/v) Tween 20). Antibody samples were diluted to 100nM and 10nM in assay buffer (PBS containing 0.5% (w/v) BSA, 0.05% (v/v) Tween 20). As a control, a reference antibody (Fab or IgG, depending on the sample format) and assay buffer were used. Samples and controls were added at 30 μ L/well and incubated for 3 hours at room temperature on a microtiter plate shaker. The plates were washed three times and 30 μ L of detection antibody (ECL-labeled anti-human Fab) was added per well and incubated for one hour on a microtiter plate shaker (approximately 500 rpm). After washing the MSD plates and adding 35. mu.L/well of MSD reading buffer T and surfactant, the electrochemiluminescence signal was detected using Sector Imager 6000(Meso Scale Discovery, Gathersburg, Md., USA). For evaluation, the signal of the antibody sample on the specific protein was divided by the corresponding signal of the reference mAb to give the Binding Ratio (BR). The Cumulative Binding Ratio (CBR) was then calculated for all proteins (25 in total) except the control: CBR up to 150 represents no detectable non-specific binding of the antibody or fragment thereof. The above values represent antibodies or fragments thereof with increased non-specific binding compared to the reference mAb.

Table 10: maturation of the plant

Summary of candidates

Mature antibodies	Parent antibody	Mature CDR
			XX01	WW01	HCDR2
XX02	WW06	HCDR2
			XX03	WW03	HCDR2
XX04	WW03	LCDR3
			XX05	WW06	HCDR2
XX06	WW03	LCDR3
			XX07	WW03	LCDR3
XX08	WW01	HCDR2
			XX10	WW06	HCDR2
XX12	WW03	LCDR3

In addition, these clones were tested for binding to the following antigens by ELISA: human NPR1, a constitutively active human NPR1 mutant (W74R), rat NPR1, human NPR3 (reverse target), each in the presence and absence of ANP and BSA. The binding of the clones to CHO K1 cells expressing human NPR1, and to the parental CHO K1 cells in the absence and presence of ANP (100nM) was also analyzed by flow cytometry. The binding characteristics of the 10 candidates are shown in fig. 9 and 10. Figure 11 shows the ANP competition results for nine of these candidates.

Negative values for XX01, XX03, XX04, XX06, XX07 and XX12 indicate that these antibodies enhance binding to ANP. The functional activity of 10 candidates was analyzed using a cellular cGMP generation assay, and the results are shown in fig. 12. From the functional data of the parental clone WW06, it can be expected that its derivatives XX02, XX05 and XX10 show weak to non-functional activity in the IgG format, but very high functionality in the monovalent FabCys format.

Through SET K_DThe measurements determined the affinity of the monovalent FabCys forms XX01-XX08, XX10 and XX 12. Affinity compared to parental clone (via

Determined in another experiment), the results are summarized in table 11. The affinity of WW01 and WW03 derivatives was improved 2300-fold for human and rat NPR1, while the affinity for the NPR1-ANP complex was only slightly improved (5-fold maximum). Their affinity for hNPR1 was between 10 and 46nM, for the hNPR1-ANP complex was 100 andbetween 300 pM. All WW01 and WW03 progeny showed rat cross-reactivity with rat/human K_DRatio of<5. The affinity of WW06 derivatives to human NPR1 and hNPR1-ANP complex is improved by 8 times to the maximum, K_DValues were between 1 and 5nM, but no binding to rat NPR1 or rat NPR1-ANP complex was observed.

Table 11: parent and mature

Affinity of candidate

Example 9: cross-cloning and PTM removal

The parent clone WW03 has a "DG" site in HCDR 2. Most WW03 derivatives (26 out of 27) were diverse in LCDR 3. Only one candidate (XX03) was diversified in HCDR2, including mutating "DG" to "DK" at amino acid position 54 of the heavy chain variable region (see, e.g., position 54 of SEQ ID NO: 122). The light chains of functional LCDR3 variegated clones were cross-cloned with the heavy chains of XX03 to engineer these clones without loss of functionality. In addition, the "DG" to "DK" mutations were inserted into the original heavy chains of several diverse derivatives of LCDR 3. Table 12 shows a summary of exemplary cross clones and D54K engineered candidates.

Table 12: overview of VL-VH Cross-clones and D54K engineered clones (WW03 derivatives)

Exemplary functional data for cross clone (XX16) versus original clone (XX06) and ANP are shown in FIG. 13. All cross clones had similar or even better functional activity than their original clones. XX16 even had a maximum cGMP concentration comparable to the natural ligand ANP in an in vitro study that analyzed their activity in hNPR1 transformed CHO cells (see fig. 14).

The specificity of the cross-clones and PTM-removed clones was tested via the 3P assay. Both D54K engineered clones XX13 and XX14 showed non-specific binding to several antigens and were therefore abolished, whereas no cross-clone showed non-specific binding. The results are shown in Table 13.

Table 13: in vitro functional data of XX15 and XX16

Example 10: crystal structure of anti-NPR 1 antibody

The crystal structure of several molecules forming complexes with hNPR1 is as follows.

For Fab03-WW03, the Fab construct of WW03 was complexed to the extracellular domain of hNPR1(C264T) at a molar ratio of 2 Fab molecules per 1 NPR1 molecule. The complex was incubated in the cold chamber for 1 hour, shaken and then loaded onto a Superdex 20016/60 column in buffer 20mM HEPES pH 7.4, 100mM NaCl. The complex protein was separated from the small aggregate peak and excess Fab and then concentrated to 19.8 mg/mL. The complex was crystallized in space group P212121 and diffracted to

The resolution of (2). The model was constructed using molecular replacement with hNPR1 structure and Fab molecules, iteratively in Coot, and refined with Buster to 21.7% Rfree.

For Fab06-WW06, the Fab construct of WW06 was complexed to the extracellular domain of hNPR1(C264T) at a molar ratio of 2 Fab molecules per 1 NPR1 molecule. The complex was incubated in the cold chamber for 1 hour, shaken and then loaded onto a Superdex 20016/60 column in buffer 20mM HEPES pH 7.4, 100mM NaCl. The complex protein was separated from the small aggregate peak and excess Fab and then concentrated to about 20.0 mg/mL. The complex was crystallized in space group P212121 and diffracted to

The resolution of (2). The model was constructed using molecular replacement with hNPR1 structure and Fab molecules, iteratively in Coot, and refined with Buster to 20.9% Rfree.

For Fab16-XX16, the Fab construct of XX16 was complexed to the extracellular domain of hNPR1(C264T) at a molar ratio of 2 Fab molecules per 1 NPR1 molecule. The complex was incubated in the cold chamber for 1 hour, shaken and then loaded onto a Superdex 20016/60 column in buffer 20mM HEPES pH 7.4, 100mM NaCl. The complex protein was separated from the small aggregate peak and excess Fab and then concentrated to about 20.0 mg/mL. The complex was crystallized in space group P212121 and diffracted to

The resolution of (2). The model was constructed using molecular replacement with hNPR1 structure and Fab molecules, iteratively in Coot, and refined with Buster to 24.4% Rfree.

The crystal structure of Fab06 complexed with hNPR1 is shown in figure 15. Figure 16 shows the conformation of hNPR1 ECD, as complexed with Fab 06; fab06 was removed from the image to more clearly reveal the conformation of hNPR1 induced by Fab binding. The structures shown in FIGS. 15 and 16 explain the differences between the Fab and IgG functional data shown in FIG. 6. The Fab C-termini are 180 degrees apart, excluding a single IgG that spans the receptor dimer. The structure of hNPR1-ECD complexed with ANP (also identified as part of this work) is shown in figure 17 (left) for comparison, while the crystal structure of Fab16 (the Fab form of the XX16 antibody) complexed with the extracellular domain of hNPR1 is shown on the right. The potency of WW06 Fab was significantly enhanced (cGMP production) when tested on CHO cells expressing hNPR 1W 74R/C232T (constitutively active mutant) compared to CHO cells expressing WT hNPR1 (fig. 18, panels a and B), suggesting that the mutation destabilizes the head-to-head conformation of NPR1 and promotes antibody binding/receptor activation.

Example 11:

phage library panning and screening

Six panning strategies were performed, which reflected the results from the initial panning

Or modifications to these strategies for specific methods of action. The panning strategy is summarized in detail in table 14. Strategies #2 and #5 with initial

The strategies performed in the activity were the same and were chosen because all five initial functional candidates were derived from these panning strategies. Strategies #3 and #4 are variations of the initial strategy with the emphasis on the alternate appearance of hNPR1 with cells expressing hNPR 1. In addition, a constitutively active mutant of NPR1 (W74R) was used as an antigen in strategies #1 and # 6. The exported bacterial lysate (BEL) of the third round of panning in the phage display vector pYPDis was used directly for primary and secondary screening.

Table 14: summary of panning strategies

The output of the third panning round was analyzed for binding to relevant antigens and cell lines. In the absence and presence of ANP, the constitutively active hNPR1 mutant (W74R), and the reverse target human hNPR3, 368 clones/subcode (4416 clones in total) were screened in an ELISA-based primary screening of human NPR 1. The primary screen yielded 810 hits which were analyzed in the secondary screen for binding to a relevant cell line (CHO-K1 cells expressing human NPR1 in the absence and presence of ANP, parental CHO-K1 cells) and rat NPR 1. A total of 380 clones from the primary and secondary screens were preferentially selected for sequencing, considering only binding to the NPR1-ANP complex, good cell binding and rat cross-reactivity. VL and VH sequencing resulted in 138 unique clones of HCDR3 with different binding properties (table 15). Six of these clones bound only in the presence of ANP.

Table 15: HCDR3 unique hit-

In the binding characteristics of

Due to the derivation from the original

The pan candidate WW06 was significantly more active in the FabCys format than in the IgG format, so the functional screening was performed in the FabCys format rather than in the IgG format.

The 138 unique clones of HCDR3 were subcloned into the FabCys form via

The process is carried out. 111 of the 138 clones were successfully converted to FabCys form and 95 clones were selected for further analysis. 92 of the 95 FabCys passed production quality control and were analyzed in detail. Then, 30 of the 92 clones with the most promising properties were selected for passage through

IgG conversion, exploratory scale expression and S-DAS. 24 of the 30 IgG passed production quality control and were analyzed in detail.

All 92 FabCys and 24 IgG were tested for binding to the relevant antigen via ELISA and flow cytometry. In addition, cloned ANP competition and functional activity was tested in a cellular cGMP production assay. Eight functional candidates were identified and analyzed for specificity in a proteomic profiling assay (3P assay) under IgG format. For comparison, one functional candidate (WW03) in the initial activity was analyzed. YY02 and YY03 showed low non-specific binding; and YY01, YY04, YY05, YY06, YY07 and WW03 showed no non-specific binding in this assay. All 92 FabCys and 24 IgG were tested for binding to the following antigens via ELISA: human NPR1, constitutively active human NPR1 mutant (W74R), rat NPR1, human NPR3 (reverse target), in the presence or absence of ANP (100nM) and unrelated antigen. Clones were also tested by flow cytometry for binding on CHO K1 cells expressing human NPR1, as well as on parental CHO K1 cells in the absence and presence of ANP. The binding properties of seven functional candidates in the IgG format are shown in fig. 19 and 20. ANP competition was tested for all 92 FabCys and 24 IgG. At 1 μ M FabCys/IgG concentration, 22 out of 92 FabCys and 12 out of 24 IgG showed significant ANP competition > 70%. The results for seven functional candidates in the IgG format are shown in figure 21. YY02, YY05, YY06 and YY07 showed clear ANP competition. YY03 and YY04 showed "negative" ANP competition in this assay, indicating stabilization of the NPR1-ANP complex.

All 92 FabCys and 24 IgG functional activities were tested in a cellular cGMP production assay using CHO-K1 cells expressing human NPR1 in the presence and absence of ANP. 8 of the 92 FabCys and 8 of the 24 iggs were functionally active and could be assigned to different modes of action. Five of the eight clones showed higher functional activity in the presence of ANP, including YY01, YY02, YY03, and YY 04. The other three clones showed an ANP independent effect in the functional assay, namely YY05, YY06 and YY 07. All eight clones were derived from panning strategies #2, #3 and #5, whereby #2 and #5 were the initial ones

Exact repetition of panning strategies #10 and #11, resulting from the initial

Identification of five functional clones active. The cGMP assay results for seven functional candidates in IgG format are shown in figure 22. As before for derivation from initial

As seen for the active candidate WW06, the functional activity of candidates YY05 and YY07 was significantly increased in the monovalent FabCys format compared to the bivalent IgG format (fig. 23).

Seven functional candidates YY01-YY07 were analyzed for their monovalent affinity to human and rat NPR1 in the absence and presence of ANP in the monovalent FabCys form. Table 16 summarizes the results of the affinity assay and epitope ranking.

Table 16: characterization in the form of monovalent FabCys-

The affinity ranged from low nM to low μ M and was largely dependent on the presence or absence of ANP. Four of the seven functional candidates (YY01, YY02, YY03 and YY04) showed significantly improved binding in the presence of ANP. YY05 and YY07 compete with ANP for binding to NPR1 and show higher affinity in the absence of ANP. The affinity of YY06 is independent of ANP. Some candidates exhibit non-specific binding to a reference flow cell, while others have such a high affinity that their K_DThe values are close to the assay limits. YY01, YY02, YY03 and YY04 share an epitope box with the epitope from primordial origin

The active WW03 is the same. YY05 and YY07 share another epitope box with those from naive

The active WW06 is the same. YY06 binds to a single epitope box.

Example 12: reformatted as IgG-

Amplification of Fab encoding inserts by Using two biotinylated primers, from the FabCys vector

The candidates were subcloned into IgG1_ LALA vector for expression in mammalian cells. The amplified product was bound to streptavidin beads, digested with restriction enzymes, and washed, releasing the purified insert into the supernatant. Inserts were cloned into recipient vectors, DNA was transformed, and individual clones were quality controlled by colony PCR and sequencing.

Selecting five

Candidates were used for affinity maturation. YY01 and YY04 stabilized the NPR1-ANP complex, YY06 acted in an ANP independent manner, while YY05 and YY07 were ANP competitive. Binding data (ELISA, flow cytometer, ANP competition), functional data (cGMP assay), affinity and epitope box are shown in table 17.

Table 17 summary of affinity, epitope and cGMP data for YY01, YY04, YY08, YY06 and YY07 in IgG format

Example 13:

generation of mature libraries

To increase affinity and biological activity and reduce non-specificity of selected antibody candidates, the LCDR3 and HCDR1/HCDR2 regions were multiplexed using

Parallel optimization of matured modules (YMM) previously used

Technical production (van den Brulle et al (2008): A novel solid phase technology for high-throughput gene synthesis [ novel solid phase technology for high throughput gene synthesis ]](ii) a Biotechniques [ biotechnological techniques ]]Page 340-343, which is hereby incorporated by reference herein).

Cloning of the maturation library was performed in a vector encoding the parental Fab fragment. Generation of mature libraries was performed against five parent antibodies (YY01, YY04, YY05, YY06, and YY 07). For library generation, all mature candidates were treated individually. The mature library was successfully cloned and the library size was 6.2X10 ⁸And 4.5x10⁹Between cfu.

To monitor cloning efficiency, the parents HCDR1/2 and LCDR3 were replaced with MBP stuffers prior to insertion of the diversified YMMs. The digested vector fragment was ligated with a 2-fold molar excess of insert fragments with diversified HCDR1/2 or LCDR 3. Electroporation of the ligation mixture into E.coli cells results>10⁸Individual colonies. Library amplification was performed as described in the literature (Tiller et al (2013): A full synthetic human Fab anti-branched on fixed VH/VL frame peptides with fast volatile biological properties [ synthetic human Fab antibody library based on fixed VH/VL framework pairs with good biophysical properties](ii) a mAbs 5(3), page 445-470, the contents of which are hereby incorporated by reference). For quality control, approximately 10-20 individual colonies per library were randomly picked and sequenced.

Example 14: panning and screening-

Nine maturation libraries were used for four different maturation panning strategies aimed at enriching progeny with improved affinity compared to parental clones. Furthermore, rat material is included where appropriate, and panning is performed under stringent conditions involving antigen concentration and washing conditions. During panning, libraries of YY01 and YY04 (LCDR 3 only) and YY05 and YY07 were pooled, while libraries of YY06 were kept separately. The panning strategy is summarized in detail in table 18. The exported bacterial lysate (BEL) after the third panning run was used directly for ELISA-based pre-screening and SET screening.

Table 18:

overview of mature panning strategy

The output of the third panning round was used for ELISA-based pre-screening to ensure that only clones binding to NPR1 and/or NPR1-ANP complex were selected for further Solution Equilibrium Titration (SET) screening. A total of 880 clones were analyzed for improved affinity to hNPR1 and/or hNPR1-ANP complex in the SET screen compared to the parental clones.

During the SET screening process 263 only improved derivatives of HCDR1//2 or LCDR3 were identified, and after sequencing and conversion to the IgG1_ LALA form, 112 unique clones were generated. The affinity of YY05 and YY07 derivatives for NPR1 was not improved, but the affinity for NPR1-ANP complex was increased by 200-fold compared to the parental clone. The affinity of the derivative of YY01 was similar to that of the parent clone YY04, and the affinity of the derivative of the parent clone YY04 was only slightly increased. The affinity of the YY06 derivative for NPR1 was increased by 4 to 40-fold and the affinity for the NPR1-ANP complex by 7 to 70-fold. See FIG. 24 (shown with affinity for hNPR 1) and FIG. 25 (shown with affinity for hNPR1-ANP complex). Affinity (K)_D[pM]) Shown on the x-axis below the parent clone names of both figures.

Example 15: maturation candidates-

Is characterized by

Of the 112 improved candidates, 95 were selected for advanced production. 77 of 95 clones passed production quality control and aimed atBinding to the relevant antigen, binding to the relevant cell line, and functional activity in cGMP production (compared to their parental clones) were characterized. After detailed characterization of IgG, 17 candidates (see table 19 for details) were selected, generated on exploratory scale IgG production, and further analyzed via 3P assay. In addition, they are converted into FabCys form to pass through SET K_DThe affinity to human and rat NPR1 was determined in the absence and presence of ANP.

Table 19: maturation of the plant

Summary of candidates

For said maturation

The affinities of 16 of the candidates (in monovalent FabCys format) were determined via SET measurements and/or via Octet. The results compared to the affinity of the parental clones are summarized in tables 20 and 21 below.

Table 20: overview of affinity data (SET) for the FabCys forms YY01, YY06, ZZ12, ZZ13, ZZ15, ZZ16 and ZZ17

Table 21: overview of affinity data (Octet) for YY01, YY06, ZZ12, ZZ13, ZZ15, ZZ16 and ZZ17 in FabCys format

Example 16: epitope determination of ANP competitive and ANP non-competitive agonist antibodies

The crystal structure of hNPR1 and XX16 Fab complexes was used to identify the XX16 Fab epitope on hNPR 1. The XX16 Fab interaction surface on hNPR1 is formed by several contiguous and non-contiguous (i.e. non-contiguous) sequences detailed in table 22. These residues form a three-dimensional conformational epitope recognized by XX16 Fab.

Epitope mapping results for XX16 Fab (ANP non-competitive) are shown in table 22. Residue hNPR1 was numbered based on SEQ ID NO:1 (P16066). Fab residues are numbered based on their linear amino acid sequence. Of the atoms of the indicated hNPR1 residue in XX16 Fab

With at least one atom inside to account for potential water-mediated interactions.

TABLE 22 epitope mapping of XX16 Fab

Key epitope residues for binding to XX16 Fab and NPR1 were identified using structural analysis and affinity maturation data to include (first order) 99-103, 105-111, 131-133, 378; and (second stage): 33-34, 76, 82, 104, 374 and 375. Amino acids 99-103 of NPR1(SEQ ID NO:1) contain both the E106 backbone (enabling D54K to achieve affinity maturation in HCDR 2) and the W106 of the NPR1 protomer from Y101 of the two-jaw HCDR 3. Some key epitope residues are shown in table 23 below. The NPR1 regions containing these key residues include N34, W76, L82, V102-A111, H131-V134 and V374-E378.

TABLE 23 Key epitope residues of XX16 Fab (ANP non-competitive)

The crystal structures of hNPR1 and WW03 Fab complexes were used to identify the WW03 Fab epitope on hNPR 1. The interaction surface of WW03 Fab on hNPR1 was formed by several contiguous and non-contiguous (i.e., non-contiguous) sequences detailed in table 24. These residues form a three-dimensional conformational epitope recognized by WW03 Fab.

Results of epitope mapping of WW03 Fab (ANP non-competitive) are shown in table 24. Residue hNPR1 was numbered based on SEQ ID NO:1 (P16066). Fab residues are numbered based on their linear amino acid sequence. Atomic of the indicated residue hNPR1 in WW03 Fab

TABLE 24 epitope mapping of WW03 Fab

Key epitope residues for WW03 Fab and NPR1 binding determined using structural analysis and affinity maturation data include the residues shown in table 25. The NPR1 region containing these key residues includes R79, L82, L99-A111, H131-V134 and V374-T375.

TABLE 25 Key epitope residues of WW03 Fab (ANP non-competitive)

The crystal structures of hNPR1 and WW06 Fab complexes were used to identify the WW06 Fab epitope on hNPR 1. The interaction surface of WW06 Fab on hNPR1 was formed by several contiguous and non-contiguous (i.e., non-contiguous) sequences detailed in table 26. These residues form a three-dimensional conformational epitope recognized by WW06 Fab.

The results of epitope mapping of WW06 Fab (ANP competitive) are shown in table 26. Residue hNPR1 was numbered based on SEQ ID NO:1 (P16066). WW06 Fab residues were numbered based on their linear amino acid sequence. Atomic of the indicated residue hNPR1 in WW06 Fab

TABLE 26 epitope mapping of WW06 Fab (ANP Competition)

Key epitope residues determined using structural analysis and affinity maturation data for WW06 and NPR1 binding include the residues shown in table 27. The NPR1 region containing these key residues includes Y188-F198, E201-R208, V215-K238 and R294-G297.

TABLE 27 Key epitope residues of WW06(ANP Competition)

Example 17: in vivo characterization of the Effect of WW06 on plasma cGMP in mice

WW06 FabCys was used in an in vivo study of hNPR1 transgenic mice to determine the effect of this antibody on plasma cGMP levels in vivo.

For the analysis of plasma cGMP samples, use was made of¹⁵N₂、¹³C cGMP is taken as an internal standard and adopts the LC-MS/MS detection method disclosed in the following documents: oeckl and Ferger, Journal of Neuroscience Methods]203(2012) 338-; and Zhang et al, J.Chromatogr B: Analyt Technol Biomed Life Sci [ J.C.: analysis techniques in biomedicine and life sciences ]2009; 877: 513-20; the contents of each of which are hereby incorporated by reference).

Plasma cGMP concentrations increased in hNPR1 Tg mice administered WW06 Fab intravenously at the 5 minute time point, with the signal returning to baseline after 3 hours. As expected and consistent with the data shown in fig. 26 and 27, T of the FabCys antibody_1/2<And (3) 30 min. Each value shown is the average of three points collected from three individual animals. Dose response data is shown in fig. 26, and PK data is shown in fig. 27.

Example 18: effect of XX16 on Heart weight and NT-proBNP of ANP KO and WT mice

XX16 was used in vivo studies to determine its effect on cardiac weight and NT-proBNP levels in ANP knock-out (KO) and Wild Type (WT) mice.

ANP knockout mice suffer from hypertension and cardiac hypertrophy (increased HW/BW ratio). NT-proBNP is a biomarker of cardiac dysfunction. In ANP knockout and wild type mice XX16 was injected subcutaneously at a dose of 0.3 or 3mg/kg once every two weeks for four weeks.

The results are shown in fig. 28. Two weeks after the second treatment, XX16 dose-dependently reduced heart weight/body weight ratio and NT-proBNP in wild type and ANP KO mice compared to vehicle treated animals.

Example 19: acute effects of XX16 on blood pressure and urinary flow Rate in hypertensive rats

XX16 was used to determine its effect on blood pressure and urinary flow rate in hypertensive rats (stroke-prone spontaneous hypertensive rats, SHRsp). Animals were administered 0.3mg/kg XX16, 1mg/kg XX16, or vehicle control intravenously (once). Blood pressure was measured using a femoral artery catheter. Measurements were taken three hours after intravenous administration and the results are shown in figure 29.

Compared to vehicle treated animals, intravenous XX16 treatment normalized mean arterial pressure and dramatically increased urinary flow rate in hypertensive rats (SHRsp).

Example 20: chronic effects of XX16 on blood pressure and plasma cGMP in Wistar-Han rats

The chronic hemodynamic effects of XX16 in telemetered implanted normal rats were evaluated. XX16 was administered subcutaneously (at doses of 0.1, 0.3, 1, 3, 10 and 30 mg/kg). The results are shown in fig. 30. Subcutaneous administration of XX16 reduced Mean Arterial Pressure (MAP) in all but the 0.1mg/kg group. There is a bottom line (about 95mmHg) for MAP reduction. A dose-dependent effect was observed on the time to return to baseline MAP; for the 30mg/kg treatment group, this effect was greater than 70 days. Plasma cGMP concentrations have an upper limit of around 90nmol/L, after which plasma cGMP gradually decreases to baseline. This effect is very similar to the blood pressure response.

Example 21: comparison of existing anti-NPR 1 antibodies with the antibodies of the present application

The WW03 antibody was compared to the antibody 5591-IgG of PCT application No. WO 2010/065293 a1 for its ability to produce cGMP in human cells expressing hNPR1 or rat cells expressing rNPR1 (both in the presence or absence of 0.075nM human or rat ANP, respectively). In the absence of ANP in both cell lines, the WW03 antibody showed excellent potentiation. Furthermore, the WW03 antibody showed excellent potentiation on in rat cells expressing rNPR1 (regardless of the presence of ANP).

In summary, previous antibodies (such as those of WO 2010/065293, including 5591-IgG) did not exhibit activity in vivo (e.g., activation was assayed by using a cGMP assay). In contrast, the antibodies of the present application exhibit in vivo activity in both mice and rats. Furthermore, crystal structure data have been used to demonstrate that epitope binding of the antibodies described herein is different from that of the antibody of WO 2010/065293. The activity, cross-reactivity, and crystallographic data described herein demonstrate that the antibodies described in the present application have different and superior effects compared to previous antibodies.

Claims

1. An isolated antibody or antigen-binding fragment that (i) binds to natriuretic peptide receptor 1(NPR 1); and (ii) is capable of activating NPR1 in the absence of Atrial Natriuretic Peptide (ANP).

2. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of claim 1, which is ANP non-competitive.

3. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of claim 1, which is ANP competitive.

4. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of claim 1 or 2, wherein the antibody or antigen-binding fragment thereof binds to an epitope within amino acids 99-133 of SEQ ID No. 1.

5. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any of claims 1, 2 or 4, wherein the antibody or antigen-binding fragment thereof binds to a conformational epitope of human NPR1, and wherein the conformational epitope comprises at least one amino acid residue of each of (i) amino acids 99-103 of SEQ ID NO:1, (ii) amino acids 105-111 of SEQ ID NO:1, (iii) amino acids 131-134 of SEQ ID NO:1, and further binds to amino acids 375 and/or 378 of SEQ ID NO: 1.

6. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of claim 1 or 3, wherein the antibody or antigen-binding fragment thereof binds to an epitope within amino acids 188-219 of SEQ ID NO: 1.

7. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the isolated antibody or antigen-binding fragment of any of claims 1, 3 or 6, wherein the antibody or antigen-binding fragment thereof binds to a conformational epitope within NPR1, and wherein the conformational epitope comprises at least one amino acid residue of each of (i) amino acids 188-198 of SEQ ID NO:1, (ii) amino acids 201-208 of SEQ ID NO:1, (iii) amino acids 215-238 of SEQ ID NO:1, and (iv) amino acids 294-297 of SEQ ID NO:1, optionally wherein the antibody or antigen-binding fragment thereof binds to at least amino acids 188, 192, 194, 197, 201, 208, and 219 of SEQ ID NO: 1.

8. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of claims 1, 2, 4, or 5, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), and wherein:

(a) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 28; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆YADSVKG (SEQ ID NO:429) or consists of the amino acid sequence shown in YADSVKG (SEQ ID NO:429), wherein X ₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X₆Is Y or F; HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30; LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41; LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 42; and LCDR3 contains, for example, Y₁QY₂Y₃Y₄Y₅An amino acid sequence shown by PRT (SEQ ID NO:430) or consists of the amino acid sequence; wherein Y is₁Is M or Q, Y₂Is S, E, T or I, Y₃Is Y or W, Y₄Is E, V, R, A, T or M, and Y₅Is K, V, R or A;

(II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 31; HCDR2 contains, for example, X₁IX₂SX₃GX₄YX₅X₆YADSVKG (SEQ ID NO:429) or consists of the amino acid sequence shown in YADSVKG (SEQ ID NO:429), wherein X₁Is A or V, X₂Is S or E, X₃Is D or K, X₄Is S or N, X₅Is I or T, and X₆Is Y or F; the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 42, and the LCDR3 comprises or consists of Y₁QY₂Y₃Y₄Y₅An amino acid sequence shown by PRT (SEQ ID NO:430) or consists of the amino acid sequence; wherein Y is₁Is M or Q, Y₂Is S, E, T or I, Y₃Is Y or W, Y ₄Is E, V, R, A, T or M, and Y₅Is K, V, R or A;

(IV) HCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:34 and HCDR2 comprises or consists of the amino acid sequence shown in IX₁SX₂GX₃YX₄(SEQ ID NO:433) or consists of the amino acid sequence shown in (wherein X is₁Is S or E, X₂Is D or K, X₃Is S or N, and X₄Is I or T, HCDR3 comprises ammonia as shown in SEQ ID NO:36Amino acid sequence or consisting thereof, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 47, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 45, and LCDR3 comprises or consists of Y ₁QY₂Y₃Y₄Y₅An amino acid sequence shown by PRT (SEQ ID NO:430) or consists of the amino acid sequence; wherein Y is₁Is M or Q, Y₂Is S, E, T or I, Y₃Is Y or W, Y₄Is E, V, R, A, T or M, and Y₅Is K, V, R or A;

(III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:32, HCDR2 comprises₁SX₂GX₃Y (SEQ ID NO:431) or an amino acid sequence consisting thereof, wherein X₁Is S or E, X₂Is D or K, or X₃Is S or N, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 44, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 45, and the LCDR3 comprises or consists of the amino acid sequence shown as Y₁WY₂Y₃PR (SEQ ID NO:435) or consists of the amino acid sequence; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A; or

(c) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 28; HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 119; HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30; LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41; LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 42; and LCDR3 comprises e.g. QQY₁WY₂Y₃Amino group represented by PRT (SEQ ID NO:434)A sequence or consisting thereof; wherein Y is₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A;

(II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO: 31; HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO: 119; the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 30, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 41, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 42, and the LCDR3 comprises or consists of the amino acid sequence shown as QQY₁WY₂Y₃An amino acid sequence shown by PRT (SEQ ID NO:434) or consists of the amino acid sequence; wherein Y is ₁Is S, E, T or I, Y₂Is V, R, A, T or M, and Y₃Is K, V, R or A;

(d) (I) HCDR1 comprises e.g. GFTFX₁An amino acid sequence represented by THYIH (SEQ ID NO:436) or a combination thereofWherein X is₁Is N, S or Q, HCDR2 comprises e.g. SIY₁Y₂Y₃GY₄Y₅TY₆YADSVKG (SEQ ID NO:437) or an amino acid sequence consisting of the same, wherein Y₁Is S or G, Y₂Is S or G, Y₃Is S or Q, Y₄Is S, Q or G, Y₅Is S, N or M, and Y₆Is Y or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO. 17, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO. 18, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO. 19;

(III) HCDR1 comprises e.g. GFTFX₁TH (SEQ ID NO:438) or an amino acid sequence consisting thereof, wherein X ₁Is N, S or Q, HCDR2 contains e.g. SY₁SGY₂Y₃(SEQ ID NO:443) or an amino acid sequence consisting of the same, wherein Y is₁Is S or G, Y₂Is S or Q, and Y₃Is S or N, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 6, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID No. 20, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 21, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 22; or

(IV) HCDR1 contains, for example, GFX₁FSX₂YX₃(SEQ ID NO:450) or consists of an amino acid sequence shown in (SEQ ID NO:450), wherein X₁Is S or T, X₂Is S, K or R, and X₃Is W or Y, HCDR2 comprises, e.g., IY₁QY₂Y₃Y₄EY₅(SEQ ID NO:451) or consists of an amino acid sequence shown in (SEQ ID NO:451), wherein Y₁Is K or H, Y₂Is S, Q or H, Y₃Is a group of the formulae G or A,Y₄is S, H or L, and Y₅Is T or K, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:232, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:243, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239;

(III)HCDR1 contains, for example, GFX₁FSX₂Y (SEQ ID NO:448) or an amino acid sequence consisting of the same, wherein X₁Is S or T, and X₂Is S, K or R, HCDR2 comprises e.g. HQY₁Y₂Y₃E (SEQ ID NO:456) or an amino acid sequence consisting of the same, wherein Y₁Is Q or H, Y₂Is G or A, and Y₃Is H or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or

(h) (I) HCDR1 comprises, for example, GFTFSX₁YX₂IX₃(SEQ ID NO:452) or consists of an amino acid sequence as shown in (SEQ ID NO:452), wherein X₁Is S or R, X₂Is W or Y, and X₃Is S or N, HCDR2 comprises, e.g., Y₁IY₂QY₃Y₄Y₅EY₆Y₇YVESVKG (SEQ ID NO:446) or a sequence consisting of the same, wherein Y is₁Is S or N, Y₂Is K or H, Y₃Is S, Q or H, Y₄Is G or A, Y₅Is S, H or L, Y₆Is T or K, and Y₇Y, K or R, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprisesThe amino acid sequence depicted by EQ ID NO:237 or consists thereof, LCDR2 comprises or consists of the amino acid sequence depicted by SEQ ID NO:238, and LCDR3 comprises or consists of the amino acid sequence depicted by SEQ ID NO: 239;

(IV) HCDR1 comprises, for example, GFTFSX₁YX₂(SEQ ID NO:457) or an amino acid sequence consisting of the amino acid sequence, wherein X₁Is S or R, and X₂Is W or Y, HCDR2 comprises, e.g., IY₁QY₂Y₃Y₄EY₅(SEQ ID NO:451) or consists of an amino acid sequence shown in (SEQ ID NO:451), wherein Y₁Is K or H, Y₂Is S, Q or H, Y₃Is G or A, Y₄Is S, H or L, and Y₅Is T or K, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:232, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:243, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 239; or

(III) HCDR1 comprises, for example, GFTFSX₁Y(SEQ ID455) or consists thereof, wherein X₁Is S or R, HCDR2 comprises e.g. HQY1Y₂Y₃E (SEQ ID NO:456) or an amino acid sequence consisting of the same, wherein Y₁Is Q or H, Y₂Is G or A, and Y₃Is H or L, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:228, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:240, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:241, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 242; or

9. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of claims 1, 3, 6, or 7, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), and wherein:

(a) (I) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:310, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of GX₁X₂X₃GX₄LGFDH (SEQ ID NO:459) or consists of the amino acid sequence shown in the specification, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises320, LCDR2 comprises, for example, GNSNRPY₁(SEQ ID NO:460) or consists of an amino acid sequence shown in (SEQ ID NO:460) wherein Y₁Is S or N, and LCDR3 comprises, for example, QSYZ₁Z₂Z₃Z₄Z₅Z₆Z₇V (SEQ ID NO:461) wherein Z is₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V;

(II) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:229, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:311, HCDR3 comprises or consists of GX₁X₂X₃GX₄LGFDH (SEQ ID NO:459) or consists of the amino acid sequence shown in the specification, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown in SEQ ID NO:320, LCDR2 comprises, for example, GNSNRPY₁(SEQ ID NO:460) or consists of an amino acid sequence shown in (SEQ ID NO:460) wherein Y ₁Is S or N, and LCDR3 comprises, for example, QSYZ₁Z₂Z₃Z₄Z₅Z₆Z₇V (SEQ ID NO:461) wherein Z is₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V;

(III) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:80, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:313, HCDR3 comprises or consists of GX₁X₂X₃GX₄LGFDH (SEQ ID NO:459) or consists of the amino acid sequence shown in the specification, wherein X₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:323, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, andand LCDR3 comprises the same as YZ₁Z₂Z₃Z₄Z₅Z₆Z₇(SEQ ID NO:462) or an amino acid sequence consisting of the amino acid sequence shown in (SEQ ID NO:462), wherein Z₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V; or

(IV) HCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:82, HCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:314, HCDR3 comprises or consists of ARGX₁X₂X₃GX₄LGFDH (SEQ ID NO:463) or an amino acid sequence consisting of ₁Is A or S, X₂Is V or L, X₃Is A or P, and X₄Is Q or L, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:326, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:324, and LCDR3 comprises or consists of the amino acid sequence shown as QSYZ₁Z₂Z₃Z₄Z₅Z₆Z₇V (SEQ ID NO:461) wherein Z is₁Is Y, D or G, Z₂Is T, S or A, Z₃Is S, P or F, Z₄Is S, T or P, Z₅Is H, S or R, Z₆Is G, S or F, and Z₇Is P, S or V;

(b) (I) HCDR1 comprises e.g. GFTFX₁X₂YAX₃X₄(SEQ ID NO:464) or consists of an amino acid sequence as shown in (SEQ ID NO:464), wherein X₁Is S or G, X₂Is S or T, X₃Is I or M, and X₄Is S or T, HCDR2 comprises e.g. Y₁ISY₂Y₃GY₄Y₅Y₆Y₇YAY₈An amino acid sequence as shown in SVKG (SEQ ID NO:465) or consists thereof, wherein Y₁Is A or S, Y₂Is A, S or G, Y₃Is S or H, Y₄Is G or Y, Y₅Is S or Y, Y₆Is T or A, Y₇Is Y, R or N, and Y₈Is E or G, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, LCDR1 comprises the amino acid sequence shown as SEQ ID NO:337, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID No. 338 and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID No. 339;

(II) HCDR1 contains as X ₁YAX₂X₃(SEQ ID NO:466) or consists of an amino acid sequence shown in (SEQ ID NO:466) wherein X₁Is S or T, X₂Is I or M, and X₃Is S or T, HCDR2 comprises e.g. Y₁ISY₂Y₃GY₄Y₅Y₆Y₇YAY₈An amino acid sequence as shown in SVKG (SEQ ID NO:465) or consists thereof, wherein Y₁Is A or S, Y₂Is A, S or G, Y₃Is S or H, Y₄Is G or Y, Y₅Is S or Y, Y₆Is T or A, Y₇Is Y, R or N, and Y₈Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339;

(III) HCDR1 comprises e.g. GFTFX₁X₂Y (SEQ ID NO:467), wherein X is₁Is S or G, and X₂Is S or T, HCDR2 comprises e.g. SY₁Y₂GY₃Y₄(SEQ ID NO:468) or a sequence consisting of the amino acids shown in (SEQ ID NO: Y)₁Is A, S or G, Y₂Is S or H, Y₃Is G or Y, and Y₄Is S or Y, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:340, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 342; or

(IV) HCDR1 contains e.g. GFTFX₁X₂YA (SEQ ID NO:469) or an amino acid sequence consisting of the same, wherein X₁Is S or G, and X₂Is S or T, HCDR2 comprises, e.g., ISY₁Y₂GY₃Y₄T(SE470) or consists of, wherein Y is₁Is S or G, Y₂Is S or H, Y₃Is G or Y, and Y₄Is S or Y, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:332, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:343, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339; or

(c) (I) HCDR1 comprises e.g. GFTFX₁X₂YAX₃X₄(SEQ ID NO:464) or consists of an amino acid sequence as shown in (SEQ ID NO:464), wherein X₁Is S or G, X₂Is S or T, X₃Is I or M, and X₄Is S or T, HCDR2 comprises e.g. SISY₁Y₂GYYY₃Y₄YAY₅An amino acid sequence as shown in SVKG (SEQ ID NO:471) or consists thereof, wherein Y₁Is A or S, Y₂Is S or H, Y₃Is T or A, Y₄Is R or N, and Y₅Is E or G, the HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, the LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:337, the LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:338 and the LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 339;

(III) HCDR1 comprises e.g. GFTFX₁X₂Y (SEQ ID NO:467), wherein X is₁Is S or G, and X₂Is S or T, HCDR2 comprises e.g. SY₁Y₂GYY (SEQ ID NO:472) or an amino acid sequence consisting of the same, wherein Y is₁Is A or S, and Y₂Is S or H, HCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO:331, LCDR1 comprises or consists of the amino acid sequence shown as SEQ ID NO:340, LCDR2 comprises or consists of the amino acid sequence shown as SEQ ID NO:341, and LCDR3 comprises or consists of the amino acid sequence shown as SEQ ID NO: 342; or

10. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of claims 1, 2, 4, 5, or 8, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), and wherein:

11. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of claims 1, 3, 6, 7, or 9, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3), and wherein:

12. The antibody or antigen-binding fragment of any one of claims 1, 2, 4, 5, 8, or 10, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from the group consisting of seq id no:

13. The antibody or antigen-binding fragment of any one of claims 1, 3, 6, 7, 9, or 11, wherein the antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (HCDR1, HCDR2, and HCDR3) and three light chain complementarity determining regions (LCDR1, LCDR2, and LCDR3) selected from the group consisting of seq id no:

14. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of claims 1, 2, 4, 5, 8, 10, or 12, wherein the antibody or antigen-binding fragment comprises:

15. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of claims 1, 3, 6, 7, 9, 11, or 13, wherein the antibody or antigen-binding fragment comprises:

(d) 392 and 344;

16. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of claims 1, 2, 4, 5, 8, 10, 12, or 14, wherein the antibody or antigen-binding fragment comprises:

17. An isolated anti-NPR 1 antibody or antigen-binding fragment; or the antibody or antigen-binding fragment of any one of claims 1, 3, 6, 7, 9, 11, 13, or 15, wherein the antibody or antigen-binding fragment comprises:

18. The antibody or antigen-binding fragment of any one of claims 1-17, which is an antigen-binding fragment selected from the group consisting of: fab, Fab ', F (ab')₂Fv, single domain antibodies (dAbs), and single chain variable fragments (scFv).

19. The antibody or antigen-binding fragment of any one of claims 1-19, wherein the antibody or antigen-binding fragment is therapeutic.

20. One or more isolated nucleic acids encoding the amino acid sequence of the antibody or antigen-binding fragment of any one of claims 1-19.

21. A vector comprising one or more isolated nucleic acids of claim 20.

22. A host cell comprising one or more isolated nucleic acids of claim 20 or the vector of claim 21.

23. A method of producing the antibody or antigen-binding fragment of any one of claims 1-19, the method comprising culturing the host cell of claim 22 under conditions suitable for production of the antibody or antigen-binding fragment, optionally wherein the method additionally comprises purifying the antibody or antigen-binding fragment.

24. A pharmaceutical composition comprising the antibody or antigen-binding fragment of any one of claims 1-19 or produced by the method of claim 23, and a pharmaceutically acceptable carrier; optionally, wherein the composition further comprises an additional therapeutic agent.

25. The pharmaceutical composition of claim 24, or a combination comprising the antibody or antigen-binding fragment of any one of claims 1-19, wherein the additional therapeutic agent is selected from ACE (angiotensin converting enzyme) inhibitors, Angiotensin Receptor Blockers (ARBs), enkephalinase inhibitors, beta blockers, diuretics, calcium channel blockers, cardiac glycosides, sodium-glucose cotransporter 2 inhibitors (SGLT2i), angiotensin receptor-enkephalinase inhibitors (ARNi), corticosteroids, leukotriene modifiers, bronchodilators, beta-adrenoreceptor antagonists, carbonic anhydrase inhibitors, alpha 2-adrenoreceptor agonists, parasympathomimetics, prostaglandin analogs, rho kinase inhibitors, and combinations thereof.

26. The antibody or antigen-binding fragment of any one of claims 1-19, or the pharmaceutical composition of claim 24 or 25, for use in treating a disorder or disease associated with natriuretic peptide receptor activity in a subject in need of such treatment.

27. The antibody or antigen-binding fragment of any one of claims 1-19, or the pharmaceutical composition or combination of claim 24 or 25, for use in treating a cardiovascular disorder in a subject in need of such treatment, optionally wherein the cardiovascular disorder is selected from: hypertension, peripheral vascular disease, heart failure, Coronary Artery Disease (CAD), Ischemic Heart Disease (IHD), mitral stenosis and regurgitation, angina, hypertrophic cardiomyopathy, diabetic cardiomyopathy, supraventricular and ventricular arrhythmias, arrhythmia, Atrial Fibrillation (AF), new atrial fibrillation, recurrent atrial fibrillation, cardiac fibrosis, atrial flutter, detrimental vascular remodeling, plaque stabilization and Myocardial Infarction (MI).

28. The antibody or antigen-binding fragment of any one of claims 1-19, or the pharmaceutical composition or combination of claim 24 or 25, for use in treating heart failure, Hypertrophic Cardiomyopathy (HCM), hypertension, preeclampsia, asthma, glaucoma, renal disorders, and/or cytokine release syndrome in a subject in need of such treatment, optionally wherein the heart failure is selected from the group consisting of heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), heart failure after acute myocardial infarction, or acute compensatory heart failure.